Research ArticleStudy of the Binding Interaction between Wortmannin and CalfThymusDNAMultispectroscopic andMolecular Docking Studies

Shiva Mehran1 Yousef Rasmi2 Hamid Reza Karamdel2 Ramin Hossinzadeh3

and Zafar Gholinejad 4

1Department of Biology Higher Education Institute of Rabe-Rashidi Tabriz Iran2Department of Biochemistry Faculty of Medicine Urmia University of Medical Science Urmia Iran3Department of Microbiology Urmia Branch Islamic Azad University Urmia Iran4Department of Medical Laboratory Science Urmia Branch Islamic Azad University Urmia Iran

Correspondence should be addressed to Zafar Gholinejad zafargholinejad777gmailcom

Received 11 July 2019 Revised 8 September 2019 Accepted 16 October 2019 Published 28 December 2019

Guest Editor Samuel Martins Silvestre

Copyright copy 2019 ShivaMehran et al-is is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Introduction Wortmannin (WTN) is a steroid metabolite that inhibits phosphatidylinositol 3-kinase and other signalingpathways Structurally the WTN consists of a cyclopentanophenanthrene-like structure with several oxygen-rich moieties whichhave the potential to interact with deoxyribonucleic acid (DNA) molecules Methods We aim to evaluate the WTN and calfthymus DNA (ct-DNA) interaction with molecular docking using the AutoDock 42 software UV and fluorescence spectroscopyand viscosity techniques were performed to confirm the in silico analysis Results Molecular docking showed that the WTNinteracted with ct-DNA via hydrogen bonds at guanine-rich sequences -e number of hydrogen bonds between the WTN andDNA was 1-2 bonds (average 12) per WTN molecule -e in silico binding constant was 2times103Mminus 1 UV spectroscopy showedthat the WTN induced a hyperchromic feature without wavelength shifting -e WTN and DNA interaction led to quenching ofDNA-emitted fluorescence -e different concentrations of WTN had no effect on DNA viscosity Taken together our resultsdemonstrated WTN interacts with DNA in the nonintercalating mode which is considered as a new mechanism of actionConclusion -ese results suggest that the WTN may exert its biological effects at least in part via interaction with DNA

1 Introduction

In 1957 Brian et al isolated the wortmannin (WTN) fromfiltrates of Penicillium wortmannii Klocker and Millan et alproposed the molecular structure based on chemical andspectroscopic evidence which is illustrated in Figure 1 [1 2]-is steroidmetabolite contains a cyclopentanophenanthrene-like structure in which A ring is modified to lactone ring and aheterocyclic furan is added to cyclopentanophenanthrene asthe fifth ring WTN belongs to furanosteroids which originatefrom lanosterol [3]

-e subsequent studies confirmed this structure andreported several biologic functions of WTN including an-tifungal cell arresting proapoptotic and anti-inflammatoryactivities [5ndash8] Due to anti-inflammatory effects of WTN itmay be a therapeutic option to treat of inflammatory dis-orders such as gout and rheumatoid arthritis

In 1993 Yano et al reported that WTN interacts withphosphatidylinositol 3-kinase and inhibits its signalingpathway [9] -e inhibitory effects of WTN on NF-kB andMAPK signaling pathways were also reported [10ndash12]

Here we hypothesized that WTN interacts with deox-yribonucleic acid (DNA) molecules that mediate some of itsbiological function -is hypothesis is based on the evidenceindicating that most steroid metabolites modulate the DNAfunction such as gene expression by direct interaction withDNA and histones [13 14] A pioneer study by Cohen andKidson demonstrated that steroids interact with guaninehomoribopolynucleotides predominantly via hydrogen (H)bonds Polyguanine has maximum hydrogen atoms for Hbond formation and progesterone presents maximum af-finity to polynucleotides due to the oxygen-rich structure[15] On the other hand WTN contains eight oxygen atomswhich can contribute to the formation of H bonds In

HindawiEvidence-Based Complementary and Alternative MedicineVolume 2019 Article ID 4936351 7 pageshttpsdoiorg10115520194936351

addition the WTN contains a bay region at the cyclo-pentanophenanthrene-like structure that may mediate thepossible WTN-DNA interaction via the hydrophobic in-teraction [16]

-erefore here we propose that WTN may exert itsbiological effects at least in part via interaction with DNAbesides the putative inhibitory effects on signaling pathwaysNevertheless to the best of our knowledge there is no reportregarding in silico andor in vitro experiments about theWTN and DNA interaction In this study we evaluatedWTN and calf thymus DNA (ct-DNA) interaction bymolecular docking viscosity UV spectroscopy and fluo-rescence spectroscopy

2 Materials

Highly polymerized ct-DNA and WTN Penicillium funi-culosum were purchased from Sigma Chemical Co (StLouis MO USA) Tris (hydroxymethyl) aminomethanewas obtained from Merck (Darmstadt Germany) All ex-periments were performed in Tris buffer solution (10mMlsquopH 74) -e stock solution of ct-DNA was prepared bydissolving an appropriate amount of ct-DNA in buffer so-lution by shaking gently -e stock solution was stored for24 h at 4degC and utilized not more than 4 days -e con-centration of ct-DNA in stock solution was 44times10minus 4M thatdetermined by spectrophotometry at 260 nm using an ex-tinction coefficient (ε) of 6600Mminus 1middotcmminus 1 DNA purity waschecked using absorption of DNA in 260 nm (A260) and280 nm (A280) -e A260A280 ratio was 185 indicating thatthe ct-DNA was sufficiently pure and protein free [17 18]Stock solution of WTN (5times10minus 3M in ddH2O) was preparedimmediately before experiments due to instability

3 UV Absorption Spectra Analysis

UV spectra analysis is a simple technique for ligand-DNAinteraction Absorbance in 230ndash300 nm wavelength rangewas measured using a spectrophotometer (Analytikjena

spekol 2000) equipped with a 10 cm quartz cell Forquantification of the WTN-DNA binding constants theabsorption spectra measurements were performed at dif-ferent concentrations of DNA ((0 066 11 154 22)times

10minus 5M) and constant WTN concentration (1times 10minus 5M)

4 Fluorescence Emission Spectra Analysis

Fluorometric assay was performed using a ShimadzuSpectrophotometer (RF-5301 PC) equipped with a quartzcell of 10 cm path length WTN fluorescence emissionspectra were recorded at 280ndash320 nm by excitation in260 nm Analysis was performed at constant concentrationof WTN (75times10minus 6M) in presence of different ct-DNAconcentrations ((0 022lsquo 066lsquo11 154 198 242lsquo 286)times

10minus 5M)

5 Viscosity Measurements

-e effect of WTN on the viscosity of DNA was measuredusing a Lovis 2000M digital micro viscometer -e tem-perature was kept constant at 25degC -e DNA concentrationwas 66times10minus 6M while varying the WTN concentrations ((0025 05 075 1)times 10minus 5M)-e data were presented as (ηη0)13 vs [WTN][DNA] where η and η0 are the viscosity of ct-DNA in the presence and absence of WTN respectively [19]

6 Molecular Docking Study

Docking study was performed using the AutoDock Tools 42software Structure of B-DNA dodecamer (PDB ID 1BNA)was obtained from the protein data bank (httpwwwrcsborg) [20] -e WTN molecular structure was drawn andenergy was minimized using the HyperChem 806 software-e DNA file was prepared by deleting water moleculesfollowed by addition of polar H atoms and Gasteiger chargeGrid box dimensions at grid points in xtimes ytimes z directionswere set to 56lsquo 59lsquo and 49 with a grid spacing of 0375 A -ecoordinate centers of grid box were set to x 1389lsquo y 2417lsquoand z 13472 Lamarckian genetic algorithms were used tocarry out molecular docking calculations -e number ofruns was set to 100 All other parameters were assigned thedefault values [21 22]

7 Results and Discussion

71 UV Absorption Spectroscopy Figure 2(a) shows the UVspectra of the WTN-DNA complex WTN showed an ab-sorbance at 260 nm and the peak intensities were enhancedwith increasing the concentration of ct-DNA A hyper-chromic effect without red or blue shift was observed for allmolar ratios which demonstrated there is a nonintercalativebinding interaction [23 24] -e observed spectral changesproposed that WTN interacts with DNA at the groove-binding model [23] -e binding constant (Kb) was calcu-lated using equation (1) from UV data [25]

A0

A minus A0

εG

εHminus G minus εG

+εG

εHminus G minus εG

times1

Kb[DNA] (1)

O

O

O

O

O

O

O

O

H

Figure 1 WTN structure (obtained from PubChem [4])

2 Evidence-Based Complementary and Alternative Medicine

where A0 and A are the absorbance of WTN in the absenceand presence of ct-DNA respectively -e εG and εHminus G arethe absorption coefficients of WTN and WTN-DNA com-plex respectively -e Kb is 2times103Mminus 1 obtained by plottingA0A minus A0 versus 1[DNA] (Figure 2(b)) -is value for Kbsuggested the groove binding model that is similar to pre-vious studied agents (eg CundashSn2 complex 167times104Mminus 1

[26] metformin complex 83times104Mminus 1 [27] Ho(phen)2Cl3_H2O (136times104Mminus 1 [28] adefovir dipivoxil333plusmn 02times104Mminus 1 [20] and sorafenib complex56times103Mminus 1 [29])

72 Fluorescence Spectra -e fluorescence emission spectraof WTN alone and in the presence of ct-DNA are shown inFigure 3(a) WTN has emission spectra with maximumemission at about 291 nm when excited at 260 nm -efluorescence of WTN was quenched by ct-DNA without anyshift in maximum emission in a concentration-dependentmanner Reduced emission intensities (quenching) con-firmed WTN-DNA interaction Fluorescence quenching isdescribed by the SternndashVolmer equation (equation (2))[30 31]

F0

F 1 + Kqτ0[Q] 1 + Ksv[Q] (2)

where F0 and F denote the steady state fluorescence in-tensities in the absence and presence of the quencher Qrespectively Kq is the quenching rate constant of the bio-molecule τ0 is the average lifetime of the molecule withoutthe quencher [Q] is the quencher concentration and Ksv isthe SternndashVolmer quenching constant Figure 3(b) showsthe SternndashVolmer plots of F0F versus [Q]

-e calculated Ksv value was 35times105 which is higherthan the classical groove binding small molecule [32ndash34]-is severe quenching phenomenon indicated that WTNinteracts with DNA strongly and probably via the interca-lation model

Apparent binding constant (Kb) and the binding stoi-chiometry (n) were calculated according to Zhang et alrsquosequation (3) [35]

logF0 minus F( 1113857

F1113890 1113891 logKb + n log[Q] (3)

where F0 and F are the fluorescence intensity in the absenceand presence of a quencher at various concentrations [Q]respectively Figure 3(c) shows the plot of log [(F0 minus F)F]versus log [DNA] -e intercept is about 147 which in-dicates WTN binds to ct-DNA in more than one position-is observation is consistent with in silico docking analysiswhich indicates in some positionsWTN binds to ct-DNA viatwo H bonds

73 Viscosity Study Viscosity experiment is an effective toolto determine the bindingmode between small molecules andct-DNA [36] In the intercalative binding mode the DNAdouble helix separation and conformational change result inan increase in DNA viscosity while groove binding inter-action has no significant effect on the DNA viscosity [37 38]-e effect of WTN on the viscosity of ct-DNA at 25degC isshown in Figure 4 -ere is no significant change in the ct-DNA viscosity by increasing concentration of WTN -isobservation suggests that the groove binding is the mode ofinteraction between WTN and ct-DNA

74MolecularDocking Study Molecular docking study is anappealing method to understand the interaction betweenligands and DNA [39] Molecular models were made todiscuss the binding modes using the AutoDock software forthe interactions ofWTNwith ct-DNA (PDB ID 1BNA)-estructure of WTN was drawn and subjected to energy op-timization [40]100 docking runs were successfully carriedoutlsquo and the obtained runs data for the conformers are listedin Table 1 -e first conformation was taken from one of thelowest binding energy docking conformation Our resultsshowedWTN incorporated to DNA and the H bond formedbetween amino groups at C-2 on guanine and oxygen atomof the acetoxy group on C-11 In some conformations theoxygen atom of furan and lactone rings was involved in Hbond formation -ere are 63 groups in which H bondsformed including 39 groups with single H bond with

ndash0101030507091113151719

Abso

rban

ce

240 250 260 270 280 290 300230Wavelength (nm)

(a)

0005

01015

02025

03035

A 0A

ndash A

0

2 4 6 8 10 12 14 16 18 2001[ct-DNA]lowast104

(b)

Figure 2 (a) UV spectra of WTN in the presence of different concentrations of ct-DNA (pH 74 and room temperature) (b) Plot A0A minus A0versus 1[DNA] for Kb calculation

Evidence-Based Complementary and Alternative Medicine 3

0

5

10

15

20

25

30

35

40

45Fl

uore

scen

ce in

tens

ity

280 285 290 295 300 305275Wavelength (nm)

(a)

0

2

4

6

8

10

12

F 0F

05 1 15 2 25 3 350[ct-DNA]lowast10ndash5

(b)

0

02

04

06

08

1

12

Log

(F0

ndash FF)

ndash51 ndash49 ndash47 ndash45 ndash43ndash53log [DNA]

(c)

Figure 3 (a) Fluorescence emission spectra of WTN in the presence of different concentrations of ct-DNA (b) SternndashVolmer plot of F0Fversus [Q] (c) Plot of log [(F0 minus F)F] versus log [DNA]

06070809

111121314

(ηη

0)1

3

200E

ndash 01

400E

ndash 01

600E

ndash 01

800E

ndash 01

100E

+ 00

120E

+ 00

140E

+ 00

160E

+ 00

000E

+ 00

[WTN][DNA]

Figure 4 Viscosity of ct-DNA in constant concentration (66times10minus 6) at 25degC in presence of WTN at different concentrations ((0 25 5 7510)times 10minus 6)

Table 1 -e data on DNA and WTN docking obtained from the AutoDock software

Rank Run Lowest binding energy(kcalMminus 1)

Mean binding energy(kcalMminus 1)

Number ofcluster

Ki(μM)

ClusterRMSD

ReferenceRMSD

1 8 minus 683 minus 661 28 984 000 30842 63 minus 628 minus 623 29 2488 000 32523 37 minus 620 minus 608 13 2835 000 31864 31 minus 619 minus 604 4 2909 000 29515 23 minus 611 minus 601 12 3343 000 31406 4 minus 599 minus 596 3 4075 000 34947 80 minus 591 minus 586 11 4682 000 3171

4 Evidence-Based Complementary and Alternative Medicine

guanine 12 groups with two H bonds simultaneously withguanine and cytosinelsquo 11 groups with cytosine and 1 groupwith two H bonds between one WNT and two guanines-ese results proposed that WTN have interaction pre-dominantly with guanine -e energetically most desirableconformation of the docked pose is shown in Figure 5

8 Conclusion

-e results of this study indicated that WTN interacts withDNAmolecules Our results did not provide strong evidenceabout the mode of interaction but based on in silico mo-lecular docking UV absorption spectroscopy fluorescenceemission spectroscopy and viscosity measurement a fluc-tuation between intercalation and groove binding model-e H bond may be an important force in this interactionwhere DNA is a hydrogen donor and the WTN oxygen isconsidered as a H acceptor To the best of our knowledgethis is the first report that revealed a new and nontraditionalmechanism for the biologic effects of WTN

Taken together lack of a red-blue shift in UV spectra andthe value of obtained Kb it is suggested the groove-bindingmode -e UV spectra results and groove-binding modewere confirmed by in silico analysis However the high Ksvvalue suggests the intercalation mode of interaction

As a third mechanism it is possible that the interactionmay occur through both groove binding and intercalationwhich was reported for some molecules previously [41 42]

Circular dichroism isothermal titration calorimetry andionic strength and melting temperature experiments couldbe performed to elucidate the precise interaction mode

However they were not carried out due to WNT instabilityIn addition it should be determined whether the WTN-DNA interaction influences the gene expression and otherbiologic functions Taken together these results suggest thatthe WTN may exert its biological effects at least partly byinteraction with DNA besides the putative signaling pathwayinhibitory properties

Data Availability

-e data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

-e authors declare that they have no conflicts of interest

Acknowledgments

-e authors would like to thank Urmia Branch of IslamicAzad University for the finacial supports

References

[1] P W Brian P J Curtis H G Hemming and G L F NorrisldquoWortmannin an antibiotic produced by Penicillium wort-mannirdquo Transactions of the BritishMycological Society vol 40no 3 pp 365ndashIN3 1957

[2] J MacMillan A E Vanstone and S K Yeboah ldquo-estructure of wortmannin a steroidal fungal metaboliterdquoChemical Communications (London) no 11 pp 613-6141968

(a) (b)

Figure 5 (a) In silico groove binding of WTN and DNA (b) H bond (green) between WTN and guanine

Evidence-Based Complementary and Alternative Medicine 5

[3] G-Q Wang G-D Chen S-Y Qin et al ldquoBiosyntheticpathway for furanosteroid demethoxyviridin and identifica-tion of an unusual pregnane side-chain cleavagerdquo NatureCommunications vol 9 no 1 p 1838 2018

[4] National Center for Biotechnology Information PubChemDatabase Wortmannin CID312145 National Center forBiotechnology Information Bethesda MA USA 2019httpspubchemncbinlmnihgovcompoundWortmannin

[5] O Hazeki K Hazeki T Katada and M Ui ldquoInhibitory effectof wortmannin on phosphatidylinositol 3-kinase-mediatedcellular eventsrdquo Journal of Lipid Mediators and Cell Signallingvol 14 no 1ndash3 pp 259ndash261 1996

[6] V Singh V Praveen D Tripathi et al ldquoIsolation charac-terization and antifungal docking studies of wortmanninisolated from Penicillium radicumrdquo Scientific Reports vol 5no 1 p 11948 2015

[7] J Wang C Zhang P Xu Z W Yang C Z Weng andY X Lai ldquoPhosphoinositide 3-kinaseprotein kinase B reg-ulates inflammation severity via signaling of Toll-like receptor4 in severe acute pancreatitisrdquo Molecular Medicine Reportsvol 17 no 6 pp 7835ndash7844 2018

[8] J Yun Y G Lv Q Yao L Wang Y P Li and J YildquoWortmannin inhibits proliferation and induces apoptosis ofMCF-7 breast cancer cellsrdquo European Journal of Gynaeco-logical Oncology vol 33 no 4 pp 367ndash369 2012

[9] H Yano S Nakanishi K Kimura et al ldquoInhibition of his-tamine secretion by wortmannin through the blockade ofphosphatidylinositol 3-kinase in RBL-2H3 cellsrdquo Journal ofBiological Chemistry vol 268 no 34 pp 25846ndash25856 1993

[10] I M Ferby I Waga C Sakanaka K Kume and T ShimizuldquoWortmannin inhibits mitogen-activated protein kinase ac-tivation induced by platelet-activating factor in Guinea pigneutrophilsrdquo Journal of Biological Chemistry vol 269 no 48pp 30485ndash30488 1994

[11] S K Manna and B B Aggarwal ldquoWortmannin inhibits ac-tivation of nuclear transcription factors NF-κB and activatedprotein-1 induced by lipopolysaccharide and phorbol esterrdquoFEBS Letters vol 473 no 1 pp 113ndash118 2000

[12] S S Mendes A Candi M Vansteenbrugge et al ldquoMicroarrayanalyses of the effects of NF-κB or PI3K pathway inhibitors onthe LPS-induced gene expression profile in RAW2647 cellssynergistic effects of rapamycin on LPS-induced MMP9-overexpressionrdquo Cellular Signalling vol 21 no 7pp 1109ndash1122 2009

[13] K Sunaga and S S Koide ldquoInteraction of calf thymus histonesand DNA with steroidsrdquo Steroids vol 9 no 4 pp 451ndash4561967

[14] M Beato ldquoGene regulation by steroid hormonesrdquo Cellvol 56 no 3 pp 335ndash344 1989

[15] P Cohen and C Kidson ldquoInteractions of hormonal steroidswith nucleic acids I A specific requirement for guaninerdquoProceedings of the National Academy of Sciences vol 63 no 2pp 458ndash464 1969

[16] S Kashino D E Zacharias R M Peck J P GluskerT S Bhatt and M M Coombs ldquoBay region distortions incyclopenta[a]phenanthrenesrdquo Cancer Research vol 46 no 4pp 1817ndash1829 1986

[17] N Shahabadi and S Bagheri ldquoSpectroscopic and moleculardocking studies on the interaction of the drug olanzapine withcalf thymus DNArdquo Spectrochimica Acta Part AMolecular andBiomolecular Spectroscopy vol 136 pp 1454ndash1459 2015

[18] Y Zhang G Zhang P Fu Y Ma and J Zhou ldquoStudy on theinteraction of triadimenol with calf thymus DNA by multi-spectroscopic methods and molecular modelingrdquo Spectrochimica

Acta Part A Molecular and Biomolecular Spectroscopy vol 96pp 1012ndash1019 2012

[19] Y Sun S Bi D Song C Qiao D Mu and H Zhang ldquoStudyon the interaction mechanism between DNA and the mainactive components in Scutellaria baicalensis Georgirdquo Sensorsand Actuators B Chemical vol 129 no 2 pp 799ndash810 2008

[20] N Shahabadi and M Falsafi ldquoExperimental and moleculardocking studies on DNA binding interaction of adefovirdipivoxil advances toward treatment of hepatitis B virusinfectionsrdquo Spectrochimica Acta Part A Molecular and Bio-molecular Spectroscopy vol 125 pp 154ndash159 2014

[21] N Shahabadi and S Amiri ldquoSpectroscopic and computationalstudies on the interaction of DNA with pregabalin drugrdquoSpectrochimica Acta Part A Molecular and BiomolecularSpectroscopy vol 138 pp 840ndash845 2015

[22] P Zhu G Zhang Y Ma Y Zhang H Miao and Y WuldquoStudy of DNA interactions with bifenthrin by spectroscopictechniques and molecular modelingrdquo Spectrochimica ActaPart A Molecular and Biomolecular Spectroscopy vol 112pp 7ndash14 2013

[23] Z-Q Liu Y-T Li Z-Y Wu and S-F Zhangldquo[Cu4(H2O)4(dmapox)2(btc)]nmiddot10nH2O the first two-di-mensional polymeric copper(II) complex with bridgingμ-trans-oxamidate and μ4-1245-benzentetracarboxylato li-gands synthesis crystal structure and DNA binding studiesrdquoInorganica Chimica Acta vol 362 no 1 pp 71ndash77 2009

[24] D K Jangir S Charak R Mehrotra and S Kundu ldquoFTIR andcircular dichroism spectroscopic study of interaction of 5-fluorouracil with DNArdquo Journal of Photochemistry andPhotobiology B Biology vol 105 no 2 pp 143ndash148 2011

[25] G Wang C Yan D Wang D Li and Y Lu ldquoSpecific bindingof a dihydropyrimidinone derivative with DNA spectro-scopic calorimetric and modeling investigationsrdquo Journal ofLuminescence vol 132 no 7 pp 1656ndash1662 2012

[26] F Arjmand and M Muddassir ldquoDesign and synthesis ofheterobimetallic topoisomerase I and II inhibitor complexesin vitro DNA binding interaction with 5prime-GMP and 5prime-TMPand cleavage studiesrdquo Journal of Photochemistry and Photo-biology B Biology vol 101 no 1 pp 37ndash46 2010

[27] N Shahabadi and L Heidari ldquoBinding studies of the anti-diabetic drug metformin to calf thymus DNA using multi-spectroscopic methodsrdquo Spectrochimica Acta Part AMolecular and Biomolecular Spectroscopy vol 97 pp 406ndash410 2012

[28] S Niroomand M Khorasani-Motlagh M Noroozifar andA Moodi ldquoSpectroscopic studies on the binding of holmium-110-phenanthroline complex with DNArdquo Journal of Photo-chemistry and Photobiology B Biology vol 117 pp 132ndash1392012

[29] J-H Shi J Chen J Wang and Y-Y Zhu ldquoBinding inter-action between sorafenib and calf thymus DNA spectroscopicmethodology viscosity measurement and molecular dock-ingrdquo Spectrochimica Acta Part A Molecular and BiomolecularSpectroscopy vol 136 pp 443ndash450 2015

[30] J R Lakowicz and B R Masters ldquoPrinciples of fluorescencespectroscopyrdquo Journal of Biomedical Optics vol 13 no 2p 9901 2008

[31] F-L Cui J Fan J-P Li and Z-D Hu ldquoInteractions between1-benzoyl-4-p-chlorophenyl thiosemicarbazide and serumalbumin investigation by fluorescence spectroscopyrdquo Bio-organic amp Medicinal Chemistry vol 12 no 1 pp 151ndash1572004

[32] A Usman and M Ahmad ldquoBinding of bisphenol-F abisphenol analogue to calf thymus DNA by multi-

6 Evidence-Based Complementary and Alternative Medicine

0

5

10

15

20

25

30

35

40

45Fl

uore

scen

ce in

tens

ity

280 285 290 295 300 305275Wavelength (nm)

(a)

0

2

4

6

8

10

12

F 0F

05 1 15 2 25 3 350[ct-DNA]lowast10ndash5

(b)

0

02

04

06

08

1

12

Log

(F0

ndash FF)

ndash51 ndash49 ndash47 ndash45 ndash43ndash53log [DNA]

(c)

Figure 3 (a) Fluorescence emission spectra of WTN in the presence of different concentrations of ct-DNA (b) SternndashVolmer plot of F0Fversus [Q] (c) Plot of log [(F0 minus F)F] versus log [DNA]

06070809

111121314

(ηη

0)1

3

200E

ndash 01

400E

ndash 01

600E

ndash 01

800E

ndash 01

100E

+ 00

120E

+ 00

140E

+ 00

160E

+ 00

000E

+ 00

[WTN][DNA]

Figure 4 Viscosity of ct-DNA in constant concentration (66times10minus 6) at 25degC in presence of WTN at different concentrations ((0 25 5 7510)times 10minus 6)

Table 1 -e data on DNA and WTN docking obtained from the AutoDock software

Rank Run Lowest binding energy(kcalMminus 1)

Mean binding energy(kcalMminus 1)

Number ofcluster

Ki(μM)

ClusterRMSD

ReferenceRMSD

1 8 minus 683 minus 661 28 984 000 30842 63 minus 628 minus 623 29 2488 000 32523 37 minus 620 minus 608 13 2835 000 31864 31 minus 619 minus 604 4 2909 000 29515 23 minus 611 minus 601 12 3343 000 31406 4 minus 599 minus 596 3 4075 000 34947 80 minus 591 minus 586 11 4682 000 3171

4 Evidence-Based Complementary and Alternative Medicine

guanine 12 groups with two H bonds simultaneously withguanine and cytosinelsquo 11 groups with cytosine and 1 groupwith two H bonds between one WNT and two guanines-ese results proposed that WTN have interaction pre-dominantly with guanine -e energetically most desirableconformation of the docked pose is shown in Figure 5

8 Conclusion

-e results of this study indicated that WTN interacts withDNAmolecules Our results did not provide strong evidenceabout the mode of interaction but based on in silico mo-lecular docking UV absorption spectroscopy fluorescenceemission spectroscopy and viscosity measurement a fluc-tuation between intercalation and groove binding model-e H bond may be an important force in this interactionwhere DNA is a hydrogen donor and the WTN oxygen isconsidered as a H acceptor To the best of our knowledgethis is the first report that revealed a new and nontraditionalmechanism for the biologic effects of WTN

Taken together lack of a red-blue shift in UV spectra andthe value of obtained Kb it is suggested the groove-bindingmode -e UV spectra results and groove-binding modewere confirmed by in silico analysis However the high Ksvvalue suggests the intercalation mode of interaction

As a third mechanism it is possible that the interactionmay occur through both groove binding and intercalationwhich was reported for some molecules previously [41 42]

Circular dichroism isothermal titration calorimetry andionic strength and melting temperature experiments couldbe performed to elucidate the precise interaction mode

However they were not carried out due to WNT instabilityIn addition it should be determined whether the WTN-DNA interaction influences the gene expression and otherbiologic functions Taken together these results suggest thatthe WTN may exert its biological effects at least partly byinteraction with DNA besides the putative signaling pathwayinhibitory properties

Data Availability

-e data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

-e authors declare that they have no conflicts of interest

Acknowledgments

-e authors would like to thank Urmia Branch of IslamicAzad University for the finacial supports

References

[1] P W Brian P J Curtis H G Hemming and G L F NorrisldquoWortmannin an antibiotic produced by Penicillium wort-mannirdquo Transactions of the BritishMycological Society vol 40no 3 pp 365ndashIN3 1957

[2] J MacMillan A E Vanstone and S K Yeboah ldquo-estructure of wortmannin a steroidal fungal metaboliterdquoChemical Communications (London) no 11 pp 613-6141968

(a) (b)

Figure 5 (a) In silico groove binding of WTN and DNA (b) H bond (green) between WTN and guanine

Evidence-Based Complementary and Alternative Medicine 5

[3] G-Q Wang G-D Chen S-Y Qin et al ldquoBiosyntheticpathway for furanosteroid demethoxyviridin and identifica-tion of an unusual pregnane side-chain cleavagerdquo NatureCommunications vol 9 no 1 p 1838 2018

[4] National Center for Biotechnology Information PubChemDatabase Wortmannin CID312145 National Center forBiotechnology Information Bethesda MA USA 2019httpspubchemncbinlmnihgovcompoundWortmannin

[5] O Hazeki K Hazeki T Katada and M Ui ldquoInhibitory effectof wortmannin on phosphatidylinositol 3-kinase-mediatedcellular eventsrdquo Journal of Lipid Mediators and Cell Signallingvol 14 no 1ndash3 pp 259ndash261 1996

[6] V Singh V Praveen D Tripathi et al ldquoIsolation charac-terization and antifungal docking studies of wortmanninisolated from Penicillium radicumrdquo Scientific Reports vol 5no 1 p 11948 2015

[7] J Wang C Zhang P Xu Z W Yang C Z Weng andY X Lai ldquoPhosphoinositide 3-kinaseprotein kinase B reg-ulates inflammation severity via signaling of Toll-like receptor4 in severe acute pancreatitisrdquo Molecular Medicine Reportsvol 17 no 6 pp 7835ndash7844 2018

[8] J Yun Y G Lv Q Yao L Wang Y P Li and J YildquoWortmannin inhibits proliferation and induces apoptosis ofMCF-7 breast cancer cellsrdquo European Journal of Gynaeco-logical Oncology vol 33 no 4 pp 367ndash369 2012

[9] H Yano S Nakanishi K Kimura et al ldquoInhibition of his-tamine secretion by wortmannin through the blockade ofphosphatidylinositol 3-kinase in RBL-2H3 cellsrdquo Journal ofBiological Chemistry vol 268 no 34 pp 25846ndash25856 1993

[10] I M Ferby I Waga C Sakanaka K Kume and T ShimizuldquoWortmannin inhibits mitogen-activated protein kinase ac-tivation induced by platelet-activating factor in Guinea pigneutrophilsrdquo Journal of Biological Chemistry vol 269 no 48pp 30485ndash30488 1994

[11] S K Manna and B B Aggarwal ldquoWortmannin inhibits ac-tivation of nuclear transcription factors NF-κB and activatedprotein-1 induced by lipopolysaccharide and phorbol esterrdquoFEBS Letters vol 473 no 1 pp 113ndash118 2000

[12] S S Mendes A Candi M Vansteenbrugge et al ldquoMicroarrayanalyses of the effects of NF-κB or PI3K pathway inhibitors onthe LPS-induced gene expression profile in RAW2647 cellssynergistic effects of rapamycin on LPS-induced MMP9-overexpressionrdquo Cellular Signalling vol 21 no 7pp 1109ndash1122 2009

[13] K Sunaga and S S Koide ldquoInteraction of calf thymus histonesand DNA with steroidsrdquo Steroids vol 9 no 4 pp 451ndash4561967

[14] M Beato ldquoGene regulation by steroid hormonesrdquo Cellvol 56 no 3 pp 335ndash344 1989

[15] P Cohen and C Kidson ldquoInteractions of hormonal steroidswith nucleic acids I A specific requirement for guaninerdquoProceedings of the National Academy of Sciences vol 63 no 2pp 458ndash464 1969

[16] S Kashino D E Zacharias R M Peck J P GluskerT S Bhatt and M M Coombs ldquoBay region distortions incyclopenta[a]phenanthrenesrdquo Cancer Research vol 46 no 4pp 1817ndash1829 1986

[17] N Shahabadi and S Bagheri ldquoSpectroscopic and moleculardocking studies on the interaction of the drug olanzapine withcalf thymus DNArdquo Spectrochimica Acta Part AMolecular andBiomolecular Spectroscopy vol 136 pp 1454ndash1459 2015

[18] Y Zhang G Zhang P Fu Y Ma and J Zhou ldquoStudy on theinteraction of triadimenol with calf thymus DNA by multi-spectroscopic methods and molecular modelingrdquo Spectrochimica

Acta Part A Molecular and Biomolecular Spectroscopy vol 96pp 1012ndash1019 2012

[19] Y Sun S Bi D Song C Qiao D Mu and H Zhang ldquoStudyon the interaction mechanism between DNA and the mainactive components in Scutellaria baicalensis Georgirdquo Sensorsand Actuators B Chemical vol 129 no 2 pp 799ndash810 2008

[20] N Shahabadi and M Falsafi ldquoExperimental and moleculardocking studies on DNA binding interaction of adefovirdipivoxil advances toward treatment of hepatitis B virusinfectionsrdquo Spectrochimica Acta Part A Molecular and Bio-molecular Spectroscopy vol 125 pp 154ndash159 2014

[21] N Shahabadi and S Amiri ldquoSpectroscopic and computationalstudies on the interaction of DNA with pregabalin drugrdquoSpectrochimica Acta Part A Molecular and BiomolecularSpectroscopy vol 138 pp 840ndash845 2015

[22] P Zhu G Zhang Y Ma Y Zhang H Miao and Y WuldquoStudy of DNA interactions with bifenthrin by spectroscopictechniques and molecular modelingrdquo Spectrochimica ActaPart A Molecular and Biomolecular Spectroscopy vol 112pp 7ndash14 2013

[23] Z-Q Liu Y-T Li Z-Y Wu and S-F Zhangldquo[Cu4(H2O)4(dmapox)2(btc)]nmiddot10nH2O the first two-di-mensional polymeric copper(II) complex with bridgingμ-trans-oxamidate and μ4-1245-benzentetracarboxylato li-gands synthesis crystal structure and DNA binding studiesrdquoInorganica Chimica Acta vol 362 no 1 pp 71ndash77 2009

[24] D K Jangir S Charak R Mehrotra and S Kundu ldquoFTIR andcircular dichroism spectroscopic study of interaction of 5-fluorouracil with DNArdquo Journal of Photochemistry andPhotobiology B Biology vol 105 no 2 pp 143ndash148 2011

[25] G Wang C Yan D Wang D Li and Y Lu ldquoSpecific bindingof a dihydropyrimidinone derivative with DNA spectro-scopic calorimetric and modeling investigationsrdquo Journal ofLuminescence vol 132 no 7 pp 1656ndash1662 2012

[26] F Arjmand and M Muddassir ldquoDesign and synthesis ofheterobimetallic topoisomerase I and II inhibitor complexesin vitro DNA binding interaction with 5prime-GMP and 5prime-TMPand cleavage studiesrdquo Journal of Photochemistry and Photo-biology B Biology vol 101 no 1 pp 37ndash46 2010

[27] N Shahabadi and L Heidari ldquoBinding studies of the anti-diabetic drug metformin to calf thymus DNA using multi-spectroscopic methodsrdquo Spectrochimica Acta Part AMolecular and Biomolecular Spectroscopy vol 97 pp 406ndash410 2012

[28] S Niroomand M Khorasani-Motlagh M Noroozifar andA Moodi ldquoSpectroscopic studies on the binding of holmium-110-phenanthroline complex with DNArdquo Journal of Photo-chemistry and Photobiology B Biology vol 117 pp 132ndash1392012

[29] J-H Shi J Chen J Wang and Y-Y Zhu ldquoBinding inter-action between sorafenib and calf thymus DNA spectroscopicmethodology viscosity measurement and molecular dock-ingrdquo Spectrochimica Acta Part A Molecular and BiomolecularSpectroscopy vol 136 pp 443ndash450 2015

[30] J R Lakowicz and B R Masters ldquoPrinciples of fluorescencespectroscopyrdquo Journal of Biomedical Optics vol 13 no 2p 9901 2008

[31] F-L Cui J Fan J-P Li and Z-D Hu ldquoInteractions between1-benzoyl-4-p-chlorophenyl thiosemicarbazide and serumalbumin investigation by fluorescence spectroscopyrdquo Bio-organic amp Medicinal Chemistry vol 12 no 1 pp 151ndash1572004

[32] A Usman and M Ahmad ldquoBinding of bisphenol-F abisphenol analogue to calf thymus DNA by multi-

6 Evidence-Based Complementary and Alternative Medicine

guanine 12 groups with two H bonds simultaneously withguanine and cytosinelsquo 11 groups with cytosine and 1 groupwith two H bonds between one WNT and two guanines-ese results proposed that WTN have interaction pre-dominantly with guanine -e energetically most desirableconformation of the docked pose is shown in Figure 5

8 Conclusion

-e results of this study indicated that WTN interacts withDNAmolecules Our results did not provide strong evidenceabout the mode of interaction but based on in silico mo-lecular docking UV absorption spectroscopy fluorescenceemission spectroscopy and viscosity measurement a fluc-tuation between intercalation and groove binding model-e H bond may be an important force in this interactionwhere DNA is a hydrogen donor and the WTN oxygen isconsidered as a H acceptor To the best of our knowledgethis is the first report that revealed a new and nontraditionalmechanism for the biologic effects of WTN

Taken together lack of a red-blue shift in UV spectra andthe value of obtained Kb it is suggested the groove-bindingmode -e UV spectra results and groove-binding modewere confirmed by in silico analysis However the high Ksvvalue suggests the intercalation mode of interaction

As a third mechanism it is possible that the interactionmay occur through both groove binding and intercalationwhich was reported for some molecules previously [41 42]

Circular dichroism isothermal titration calorimetry andionic strength and melting temperature experiments couldbe performed to elucidate the precise interaction mode

However they were not carried out due to WNT instabilityIn addition it should be determined whether the WTN-DNA interaction influences the gene expression and otherbiologic functions Taken together these results suggest thatthe WTN may exert its biological effects at least partly byinteraction with DNA besides the putative signaling pathwayinhibitory properties

Data Availability

-e data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

-e authors declare that they have no conflicts of interest

Acknowledgments

-e authors would like to thank Urmia Branch of IslamicAzad University for the finacial supports

References

[1] P W Brian P J Curtis H G Hemming and G L F NorrisldquoWortmannin an antibiotic produced by Penicillium wort-mannirdquo Transactions of the BritishMycological Society vol 40no 3 pp 365ndashIN3 1957

[2] J MacMillan A E Vanstone and S K Yeboah ldquo-estructure of wortmannin a steroidal fungal metaboliterdquoChemical Communications (London) no 11 pp 613-6141968

(a) (b)

Figure 5 (a) In silico groove binding of WTN and DNA (b) H bond (green) between WTN and guanine

Evidence-Based Complementary and Alternative Medicine 5

[3] G-Q Wang G-D Chen S-Y Qin et al ldquoBiosyntheticpathway for furanosteroid demethoxyviridin and identifica-tion of an unusual pregnane side-chain cleavagerdquo NatureCommunications vol 9 no 1 p 1838 2018

[4] National Center for Biotechnology Information PubChemDatabase Wortmannin CID312145 National Center forBiotechnology Information Bethesda MA USA 2019httpspubchemncbinlmnihgovcompoundWortmannin

[5] O Hazeki K Hazeki T Katada and M Ui ldquoInhibitory effectof wortmannin on phosphatidylinositol 3-kinase-mediatedcellular eventsrdquo Journal of Lipid Mediators and Cell Signallingvol 14 no 1ndash3 pp 259ndash261 1996

[6] V Singh V Praveen D Tripathi et al ldquoIsolation charac-terization and antifungal docking studies of wortmanninisolated from Penicillium radicumrdquo Scientific Reports vol 5no 1 p 11948 2015

[7] J Wang C Zhang P Xu Z W Yang C Z Weng andY X Lai ldquoPhosphoinositide 3-kinaseprotein kinase B reg-ulates inflammation severity via signaling of Toll-like receptor4 in severe acute pancreatitisrdquo Molecular Medicine Reportsvol 17 no 6 pp 7835ndash7844 2018

[8] J Yun Y G Lv Q Yao L Wang Y P Li and J YildquoWortmannin inhibits proliferation and induces apoptosis ofMCF-7 breast cancer cellsrdquo European Journal of Gynaeco-logical Oncology vol 33 no 4 pp 367ndash369 2012

[9] H Yano S Nakanishi K Kimura et al ldquoInhibition of his-tamine secretion by wortmannin through the blockade ofphosphatidylinositol 3-kinase in RBL-2H3 cellsrdquo Journal ofBiological Chemistry vol 268 no 34 pp 25846ndash25856 1993

[10] I M Ferby I Waga C Sakanaka K Kume and T ShimizuldquoWortmannin inhibits mitogen-activated protein kinase ac-tivation induced by platelet-activating factor in Guinea pigneutrophilsrdquo Journal of Biological Chemistry vol 269 no 48pp 30485ndash30488 1994

[11] S K Manna and B B Aggarwal ldquoWortmannin inhibits ac-tivation of nuclear transcription factors NF-κB and activatedprotein-1 induced by lipopolysaccharide and phorbol esterrdquoFEBS Letters vol 473 no 1 pp 113ndash118 2000

[12] S S Mendes A Candi M Vansteenbrugge et al ldquoMicroarrayanalyses of the effects of NF-κB or PI3K pathway inhibitors onthe LPS-induced gene expression profile in RAW2647 cellssynergistic effects of rapamycin on LPS-induced MMP9-overexpressionrdquo Cellular Signalling vol 21 no 7pp 1109ndash1122 2009

[13] K Sunaga and S S Koide ldquoInteraction of calf thymus histonesand DNA with steroidsrdquo Steroids vol 9 no 4 pp 451ndash4561967

[14] M Beato ldquoGene regulation by steroid hormonesrdquo Cellvol 56 no 3 pp 335ndash344 1989

[15] P Cohen and C Kidson ldquoInteractions of hormonal steroidswith nucleic acids I A specific requirement for guaninerdquoProceedings of the National Academy of Sciences vol 63 no 2pp 458ndash464 1969

[16] S Kashino D E Zacharias R M Peck J P GluskerT S Bhatt and M M Coombs ldquoBay region distortions incyclopenta[a]phenanthrenesrdquo Cancer Research vol 46 no 4pp 1817ndash1829 1986

[17] N Shahabadi and S Bagheri ldquoSpectroscopic and moleculardocking studies on the interaction of the drug olanzapine withcalf thymus DNArdquo Spectrochimica Acta Part AMolecular andBiomolecular Spectroscopy vol 136 pp 1454ndash1459 2015

[18] Y Zhang G Zhang P Fu Y Ma and J Zhou ldquoStudy on theinteraction of triadimenol with calf thymus DNA by multi-spectroscopic methods and molecular modelingrdquo Spectrochimica

Acta Part A Molecular and Biomolecular Spectroscopy vol 96pp 1012ndash1019 2012

[19] Y Sun S Bi D Song C Qiao D Mu and H Zhang ldquoStudyon the interaction mechanism between DNA and the mainactive components in Scutellaria baicalensis Georgirdquo Sensorsand Actuators B Chemical vol 129 no 2 pp 799ndash810 2008

[20] N Shahabadi and M Falsafi ldquoExperimental and moleculardocking studies on DNA binding interaction of adefovirdipivoxil advances toward treatment of hepatitis B virusinfectionsrdquo Spectrochimica Acta Part A Molecular and Bio-molecular Spectroscopy vol 125 pp 154ndash159 2014

[21] N Shahabadi and S Amiri ldquoSpectroscopic and computationalstudies on the interaction of DNA with pregabalin drugrdquoSpectrochimica Acta Part A Molecular and BiomolecularSpectroscopy vol 138 pp 840ndash845 2015

[22] P Zhu G Zhang Y Ma Y Zhang H Miao and Y WuldquoStudy of DNA interactions with bifenthrin by spectroscopictechniques and molecular modelingrdquo Spectrochimica ActaPart A Molecular and Biomolecular Spectroscopy vol 112pp 7ndash14 2013

[23] Z-Q Liu Y-T Li Z-Y Wu and S-F Zhangldquo[Cu4(H2O)4(dmapox)2(btc)]nmiddot10nH2O the first two-di-mensional polymeric copper(II) complex with bridgingμ-trans-oxamidate and μ4-1245-benzentetracarboxylato li-gands synthesis crystal structure and DNA binding studiesrdquoInorganica Chimica Acta vol 362 no 1 pp 71ndash77 2009

[24] D K Jangir S Charak R Mehrotra and S Kundu ldquoFTIR andcircular dichroism spectroscopic study of interaction of 5-fluorouracil with DNArdquo Journal of Photochemistry andPhotobiology B Biology vol 105 no 2 pp 143ndash148 2011

[25] G Wang C Yan D Wang D Li and Y Lu ldquoSpecific bindingof a dihydropyrimidinone derivative with DNA spectro-scopic calorimetric and modeling investigationsrdquo Journal ofLuminescence vol 132 no 7 pp 1656ndash1662 2012

[26] F Arjmand and M Muddassir ldquoDesign and synthesis ofheterobimetallic topoisomerase I and II inhibitor complexesin vitro DNA binding interaction with 5prime-GMP and 5prime-TMPand cleavage studiesrdquo Journal of Photochemistry and Photo-biology B Biology vol 101 no 1 pp 37ndash46 2010

[27] N Shahabadi and L Heidari ldquoBinding studies of the anti-diabetic drug metformin to calf thymus DNA using multi-spectroscopic methodsrdquo Spectrochimica Acta Part AMolecular and Biomolecular Spectroscopy vol 97 pp 406ndash410 2012

[28] S Niroomand M Khorasani-Motlagh M Noroozifar andA Moodi ldquoSpectroscopic studies on the binding of holmium-110-phenanthroline complex with DNArdquo Journal of Photo-chemistry and Photobiology B Biology vol 117 pp 132ndash1392012

[29] J-H Shi J Chen J Wang and Y-Y Zhu ldquoBinding inter-action between sorafenib and calf thymus DNA spectroscopicmethodology viscosity measurement and molecular dock-ingrdquo Spectrochimica Acta Part A Molecular and BiomolecularSpectroscopy vol 136 pp 443ndash450 2015

[30] J R Lakowicz and B R Masters ldquoPrinciples of fluorescencespectroscopyrdquo Journal of Biomedical Optics vol 13 no 2p 9901 2008

[31] F-L Cui J Fan J-P Li and Z-D Hu ldquoInteractions between1-benzoyl-4-p-chlorophenyl thiosemicarbazide and serumalbumin investigation by fluorescence spectroscopyrdquo Bio-organic amp Medicinal Chemistry vol 12 no 1 pp 151ndash1572004

[32] A Usman and M Ahmad ldquoBinding of bisphenol-F abisphenol analogue to calf thymus DNA by multi-

6 Evidence-Based Complementary and Alternative Medicine

[3] G-Q Wang G-D Chen S-Y Qin et al ldquoBiosyntheticpathway for furanosteroid demethoxyviridin and identifica-tion of an unusual pregnane side-chain cleavagerdquo NatureCommunications vol 9 no 1 p 1838 2018

[4] National Center for Biotechnology Information PubChemDatabase Wortmannin CID312145 National Center forBiotechnology Information Bethesda MA USA 2019httpspubchemncbinlmnihgovcompoundWortmannin

[5] O Hazeki K Hazeki T Katada and M Ui ldquoInhibitory effectof wortmannin on phosphatidylinositol 3-kinase-mediatedcellular eventsrdquo Journal of Lipid Mediators and Cell Signallingvol 14 no 1ndash3 pp 259ndash261 1996

[6] V Singh V Praveen D Tripathi et al ldquoIsolation charac-terization and antifungal docking studies of wortmanninisolated from Penicillium radicumrdquo Scientific Reports vol 5no 1 p 11948 2015

[7] J Wang C Zhang P Xu Z W Yang C Z Weng andY X Lai ldquoPhosphoinositide 3-kinaseprotein kinase B reg-ulates inflammation severity via signaling of Toll-like receptor4 in severe acute pancreatitisrdquo Molecular Medicine Reportsvol 17 no 6 pp 7835ndash7844 2018

[8] J Yun Y G Lv Q Yao L Wang Y P Li and J YildquoWortmannin inhibits proliferation and induces apoptosis ofMCF-7 breast cancer cellsrdquo European Journal of Gynaeco-logical Oncology vol 33 no 4 pp 367ndash369 2012

[9] H Yano S Nakanishi K Kimura et al ldquoInhibition of his-tamine secretion by wortmannin through the blockade ofphosphatidylinositol 3-kinase in RBL-2H3 cellsrdquo Journal ofBiological Chemistry vol 268 no 34 pp 25846ndash25856 1993

[10] I M Ferby I Waga C Sakanaka K Kume and T ShimizuldquoWortmannin inhibits mitogen-activated protein kinase ac-tivation induced by platelet-activating factor in Guinea pigneutrophilsrdquo Journal of Biological Chemistry vol 269 no 48pp 30485ndash30488 1994

[11] S K Manna and B B Aggarwal ldquoWortmannin inhibits ac-tivation of nuclear transcription factors NF-κB and activatedprotein-1 induced by lipopolysaccharide and phorbol esterrdquoFEBS Letters vol 473 no 1 pp 113ndash118 2000

[12] S S Mendes A Candi M Vansteenbrugge et al ldquoMicroarrayanalyses of the effects of NF-κB or PI3K pathway inhibitors onthe LPS-induced gene expression profile in RAW2647 cellssynergistic effects of rapamycin on LPS-induced MMP9-overexpressionrdquo Cellular Signalling vol 21 no 7pp 1109ndash1122 2009

[13] K Sunaga and S S Koide ldquoInteraction of calf thymus histonesand DNA with steroidsrdquo Steroids vol 9 no 4 pp 451ndash4561967

[14] M Beato ldquoGene regulation by steroid hormonesrdquo Cellvol 56 no 3 pp 335ndash344 1989

[15] P Cohen and C Kidson ldquoInteractions of hormonal steroidswith nucleic acids I A specific requirement for guaninerdquoProceedings of the National Academy of Sciences vol 63 no 2pp 458ndash464 1969

[16] S Kashino D E Zacharias R M Peck J P GluskerT S Bhatt and M M Coombs ldquoBay region distortions incyclopenta[a]phenanthrenesrdquo Cancer Research vol 46 no 4pp 1817ndash1829 1986

[17] N Shahabadi and S Bagheri ldquoSpectroscopic and moleculardocking studies on the interaction of the drug olanzapine withcalf thymus DNArdquo Spectrochimica Acta Part AMolecular andBiomolecular Spectroscopy vol 136 pp 1454ndash1459 2015

[18] Y Zhang G Zhang P Fu Y Ma and J Zhou ldquoStudy on theinteraction of triadimenol with calf thymus DNA by multi-spectroscopic methods and molecular modelingrdquo Spectrochimica

Acta Part A Molecular and Biomolecular Spectroscopy vol 96pp 1012ndash1019 2012

[19] Y Sun S Bi D Song C Qiao D Mu and H Zhang ldquoStudyon the interaction mechanism between DNA and the mainactive components in Scutellaria baicalensis Georgirdquo Sensorsand Actuators B Chemical vol 129 no 2 pp 799ndash810 2008

[20] N Shahabadi and M Falsafi ldquoExperimental and moleculardocking studies on DNA binding interaction of adefovirdipivoxil advances toward treatment of hepatitis B virusinfectionsrdquo Spectrochimica Acta Part A Molecular and Bio-molecular Spectroscopy vol 125 pp 154ndash159 2014

[21] N Shahabadi and S Amiri ldquoSpectroscopic and computationalstudies on the interaction of DNA with pregabalin drugrdquoSpectrochimica Acta Part A Molecular and BiomolecularSpectroscopy vol 138 pp 840ndash845 2015

[22] P Zhu G Zhang Y Ma Y Zhang H Miao and Y WuldquoStudy of DNA interactions with bifenthrin by spectroscopictechniques and molecular modelingrdquo Spectrochimica ActaPart A Molecular and Biomolecular Spectroscopy vol 112pp 7ndash14 2013

[23] Z-Q Liu Y-T Li Z-Y Wu and S-F Zhangldquo[Cu4(H2O)4(dmapox)2(btc)]nmiddot10nH2O the first two-di-mensional polymeric copper(II) complex with bridgingμ-trans-oxamidate and μ4-1245-benzentetracarboxylato li-gands synthesis crystal structure and DNA binding studiesrdquoInorganica Chimica Acta vol 362 no 1 pp 71ndash77 2009

[24] D K Jangir S Charak R Mehrotra and S Kundu ldquoFTIR andcircular dichroism spectroscopic study of interaction of 5-fluorouracil with DNArdquo Journal of Photochemistry andPhotobiology B Biology vol 105 no 2 pp 143ndash148 2011

[25] G Wang C Yan D Wang D Li and Y Lu ldquoSpecific bindingof a dihydropyrimidinone derivative with DNA spectro-scopic calorimetric and modeling investigationsrdquo Journal ofLuminescence vol 132 no 7 pp 1656ndash1662 2012

[26] F Arjmand and M Muddassir ldquoDesign and synthesis ofheterobimetallic topoisomerase I and II inhibitor complexesin vitro DNA binding interaction with 5prime-GMP and 5prime-TMPand cleavage studiesrdquo Journal of Photochemistry and Photo-biology B Biology vol 101 no 1 pp 37ndash46 2010

[27] N Shahabadi and L Heidari ldquoBinding studies of the anti-diabetic drug metformin to calf thymus DNA using multi-spectroscopic methodsrdquo Spectrochimica Acta Part AMolecular and Biomolecular Spectroscopy vol 97 pp 406ndash410 2012

[28] S Niroomand M Khorasani-Motlagh M Noroozifar andA Moodi ldquoSpectroscopic studies on the binding of holmium-110-phenanthroline complex with DNArdquo Journal of Photo-chemistry and Photobiology B Biology vol 117 pp 132ndash1392012

[29] J-H Shi J Chen J Wang and Y-Y Zhu ldquoBinding inter-action between sorafenib and calf thymus DNA spectroscopicmethodology viscosity measurement and molecular dock-ingrdquo Spectrochimica Acta Part A Molecular and BiomolecularSpectroscopy vol 136 pp 443ndash450 2015

[30] J R Lakowicz and B R Masters ldquoPrinciples of fluorescencespectroscopyrdquo Journal of Biomedical Optics vol 13 no 2p 9901 2008

[31] F-L Cui J Fan J-P Li and Z-D Hu ldquoInteractions between1-benzoyl-4-p-chlorophenyl thiosemicarbazide and serumalbumin investigation by fluorescence spectroscopyrdquo Bio-organic amp Medicinal Chemistry vol 12 no 1 pp 151ndash1572004

[32] A Usman and M Ahmad ldquoBinding of bisphenol-F abisphenol analogue to calf thymus DNA by multi-

6 Evidence-Based Complementary and Alternative Medicine

spectroscopic and molecular docking studiesrdquo Chemospherevol 181 pp 536ndash543 2017

[33] I Ahmad and M Ahmad ldquoDacarbazine as a minor groovebinder of DNA spectroscopic biophysical and moleculardocking studiesrdquo International Journal of Biological Macro-molecules vol 79 pp 193ndash200 2015

[34] A Nowicka S Hafner and M Hepel ldquoAntineoplastic druginteractions with DNA modified gold piezoelectrodesrdquo ECSTransactions vol 19 no 28 pp 1ndash13 2009

[35] G Zhang X Hu and P Fu ldquoSpectroscopic studies on theinteraction between carbaryl and calf thymus DNA with theuse of ethidium bromide as a fluorescence proberdquo Journal ofPhotochemistry and Photobiology B Biology vol 108pp 53ndash61 2012

[36] N Shahabadi and N H Moghadam ldquoDetermining the modeof interaction of calf thymus DNA with the drug sumatriptanusing voltammetric and spectroscopic techniquesrdquo Spec-trochimica Acta Part A Molecular and Biomolecular Spec-troscopy vol 99 pp 18ndash22 2012

[37] G Zhang L Wang X Zhou Y Li and D Gong ldquoBindingcharacteristics of sodium saccharin with calf thymus DNA invitrordquo Journal of Agricultural and Food Chemistry vol 62no 4 pp 991ndash1000 2014

[38] Z-C Liu B-D Wang Z-Y Yang Y Li D-D Qin andT-R Li ldquoSynthesis crystal structure DNA interaction andantioxidant activities of two novel water-soluble Cu(2+)complexes derivated from 2-oxo-quinoline-3-carbaldehydeSchiff-basesrdquo European Journal of Medicinal Chemistryvol 44 no 11 pp 4477ndash4484 2009

[39] I Haq and J Ladbury ldquoDrug-DNA recognition energeticsand implications for designrdquo Journal of Molecular Recogni-tion vol 13 no 4 pp 188ndash197 2000

[40] E M Proudfoot J P Mackay and P Karuso ldquoProbing sitespecificity of DNA binding metallointercalators by NMRspectroscopy and molecular modelingrdquo Biochemistry vol 40no 15 pp 4867ndash4878 2001

[41] D Banerjee and S K Pal ldquoSimultaneous binding of minorgroove binder and intercalator to dodecamer DNA impor-tance of relative orientation of donor and acceptor in FRETrdquoCe Journal of Physical Chemistry B vol 111 no 19pp 5047ndash5052 2007

[42] E Trotta E DrsquoAmbrosio G Ravagnan and M Paci ldquoSi-multaneous and different binding mechanisms of 4prime6-Dia-midino-2-phenylindole to DNA hexamer (d(CGATCG))2a1H NMR studyrdquo Journal of Biological Chemistry vol 271no 44 pp 27608ndash27614 1996

Evidence-Based Complementary and Alternative Medicine 7

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Computational and Mathematical Methods in Medicine

Hindawiwwwhindawicom Volume 2018

Behavioural Neurology

OphthalmologyJournal of

Hindawiwwwhindawicom Volume 2018

Diabetes ResearchJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Research and TreatmentAIDS

Hindawiwwwhindawicom Volume 2018

Gastroenterology Research and Practice

Hindawiwwwhindawicom Volume 2018

Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Computational and Mathematical Methods in Medicine

Hindawiwwwhindawicom Volume 2018

Behavioural Neurology

OphthalmologyJournal of

Hindawiwwwhindawicom Volume 2018

Diabetes ResearchJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Research and TreatmentAIDS

Hindawiwwwhindawicom Volume 2018

Gastroenterology Research and Practice

Hindawiwwwhindawicom Volume 2018

Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom