clinicalsignificanceoftheinterleukin24mrnalevelinheadand ...2020/02/23 · and its subgroups to a...

Research ArticleClinicalSignificanceof the Interleukin24mRNALevel inHeadandNeck Squamous Cell Carcinoma and Its Subgroups: An InSilico Investigation

Lan-Lan Qiu,1,2 Xiao-Guohui Zhang,1 Gang Chen ,3 Yi-Wu Dang,3 Zhi-Guang Huang,3

Ming-Xuan Li,3 Yao Liang,1 Su-Ning Huang,4 Xiao-Zhun Tang,5 Xiao-Xia Chen,1

Hong-Yu Wei ,6 and Hua-Yu Wu 1

1Department of Cell Biology and Genetics, School of Pre-clinical Medicine, Guangxi Medical University, Nanning 530021,Guangxi Zhuang Autonomous Region, China2Department of Pathophysiology, School of Pre-clinical Medicine, Guangxi Medical University, Nanning 530021,Guangxi Zhuang Autonomous Region, China3Department of Pathology, First Affiliated Hospital, Guangxi Medical University, Nanning 530021,Guangxi Zhuang Autonomous Region, China4Department of Head and Neck Tumor Surgery, Guangxi Medical University Cancer Hospital, Nanning 530021,Guangxi Zhuang Autonomous Region, China5Department of Radiotherapy, Guangxi Medical University Cancer Hospital, Nanning 530021,Guangxi Zhuang Autonomous Region, China6Department of Organic Chemistry and Medicinal Chemistry, Pharmaceutical College, Guangxi Medical University,Nanning 530021, Guangxi Zhuang Autonomous Region, China

Correspondence should be addressed to Hong-Yu Wei; [email protected] and Hua-Yu Wu; [email protected]

Received 23 February 2020; Accepted 2 July 2020; Published 18 September 2020

Academic Editor: Ferdinand Frauscher

Copyright © 2020 Lan-Lan Qiu et al. /is is an open access article distributed under the Creative Commons Attribution License,which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

IL24 mRNA is known to have an apoptotic effect on cancer cells but not on noncancer cells. However, the expression level of theIL24 mRNA in head and neck squamous cell carcinoma (HNSCC) and its subgroups is rarely studied. In this study, the clinicalimplication of IL24 mRNAwas evaluated in the common subgroups of HNSCC, including oral squamous cell carcinoma (OSCC),nasopharyngeal carcinoma (NPC), and laryngeal squamous cell carcinoma (LSCC) for analysis. Substantial IL24 mRNA ex-pression data were calculated from several databases, such as the Gene Expression Omnibus (GEO), ArrayExpress, Sequence ReadArchive (SRA), ONCOMINE, and/e Cancer Genome Atlas (TCGA) databases. We ultimately collected a total of 41 microarraysand RNA-seq including 1,564 HNSCC and 603 noncancer tissue samples. IL24 mRNA was highly expressed in OSCC, LSCC, andNPC as shown by the separated standard mean difference (SMD), as well as HNSCC as a whole part (SMD� 1.47, 95% confdenceinterval (CI)� 1.24−1.70, P< 0.0001). In all subgroups, the IL24 mRNA upregulation had the ability to distinguish cancer fromnoncancer tissue with area under the curves (AUCs) of the summary receiver operating characteristic (sROC) higher than 0.85. Inconclusion, IL24 mRNA may be used as a potential marker for cancer screening, and its clinical diagnostic value needs to befurther studied. It also provides a new idea for the treatment of the IL24 gene in HNSCC and its subgroups in the future.

1. Introduction

Head and neck squamous cell carcinoma (HNSCC) is thesixth most common cancer in the world and has a five-yearsurvival rate of less than 50% [1, 2]. HNSCC has high

metastasis and recurrence rates and includes the followingsubgroups: oral squamous cell carcinoma (OSCC), naso-pharyngeal carcinoma (NPC), and laryngeal squamous cellcarcinoma (LSCC) [2, 3]. Similar to other cancers, HNSCC iscaused by abnormal genetic changes, such as point

HindawiJournal of OncologyVolume 2020, Article ID 7042025, 15 pageshttps://doi.org/10.1155/2020/7042025

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mutations, amplifications, gene rearrangements, and dele-tions, which can promote the development of tumors [4].HNSCC is also associated with a variety of environmentalfactors as known risk factors, including smoking, alcoholabuse, and human papillomavirus (HPV) infection [5]. /esurvival rate of patients with the disease has increased due toprogress in surgical therapy, as well as radiotherapy andchemotherapy. In addition, immunotherapy has become apromising treatment for HNSCC [6, 7]. However, due to thelack of obvious early clinical symptoms, many HNSCCpatients are diagnosed with advanced cancer. Moreover, theprognosis of HNSCC patients remains stagnant, with aconsiderable number of deaths due to recurrence and me-tastasis after chemotherapy and targeted therapy [8]. Fur-thermore, there are no suitable tumor markers for earlydiagnosis and prognosis evaluation of HNSCC [4].

/e Interleukin 24 mRNA (IL24 mRNA), a single copygene, is located at chromosome 1q32-41. IL24 is expressed as acytokine in a variety of immune tissues and cells at normalphysiological levels. At the supraphysiological level, IL24mRNA significantly inhibits tumor growth, invasion, metas-tasis, and angiogenesis [9]. It is a polyergic immunoregulatorycytokine that belongs to the IL-10 gene family, with a widerange of antitumor properties, including specific induction oftumor cell apoptosis, inhibition of tumor angiogenesis, andregulation of antitumor immune responses [10, 11]. Otherstudies have shown that IL24 mRNA can enhance the im-munogenicity of tumor cells through upregulation of cos-timulatory molecules, such as CD80 and CD86 [12]. Somestudies show that IL24 mRNA can inhibit several types ofcancer cells, such as prostate cancer, kidney cancer, osteo-sarcoma, melanoma, colon cancer, cervical cancer, malignantglioma, breast cancer, and lung cancer, without obvious ad-verse effects on normal cells [13]. Recent evidence indicates thatthe IL24 mRNA is an underlying candidate for gene treatmentof cancer through boosting apoptosis in many types of cancercells [14–17]. IL24 has been reported to play vital roles in theinhibition of tumor growth and induction of tumor cell ap-optosis in HNSCC cells [18, 19]. Adenovirus-mediated IL24mRNA therapy could also enhance the in vivo and in vitroantitumor effects of radiotherapy for NPC [20, 21].

However, the literature retrieved above only discussedthe treatment of HNSCC and its subgroups with IL24mRNA in cell lines or animals, and the current studies havenot further analyzed the safety assessment and clinicaltreatment trials as a feasible means. /e expression of theIL24 mRNA in HNSCC and its subgroups and the ability ofthe IL24 mRNA to distinguish cancer tissue from noncancertissue are rarely studied. /erefore, we collected 41 data setsof microarrays and RNA-Seq from several databases andspeculated that there may be a close relationship betweenIL24 mRNA expression and HNSCC. In this study, weanalyzed IL24 mRNA expression in HNSCC and its sub-groups, OSCC, LSCC, andNPC, as well as the correspondingclinical parameters. /e results showed that a high ex-pression level of the IL24 mRNA correlated with HNSCCand its subgroups to a certain extent, which potentially hasdiagnostic ability and therapeutic significance and could befurther studied as a potential factor in the future.

2. Materials and Methods

2.1.Mining the RNA-Seq Data of the IL24mRNABased on theCancer Genome Atlas (TCGA) Database. After mining theTCGA database, we downloaded the RNA-Seq data ofmRNA expression and clinicopathological parameters inHNSCC and extracted data on the IL24 mRNA, whichinvolved 502 cases with HNSCC samples (including 40 caseswith tonsil samples, 341 cases with oral cavity samples, 10cases with hypopharynx samples, and 111 cases with larynxsamples) and 44 cases with noncancer samples as a controlgroup. /e extracted data were used to analyze the ex-pression and clinical parameters, such as gender, age, andpathological stage, of patients with the IL24 mRNA inHNSCC and its subgroups. /e prognostic value of IL24mRNA was estimated by using the Kaplan–Meier survivalanalysis. Figure 1 (in the supplementary materials) shows thedata screening and flow chart of the study.

2.2. Mining the Microarrays of the IL24 mRNA Based on theGene Expression Omnibus (GEO) and ArrayExpressDatabases. After mining the data from GEO, ArrayExpress,Sequence Read Archive (SRA), and Oncomine databases, wedownloaded the microarray data of IL24 mRNA expressionin HNSCC and its subgroups and screened the data thatconformed to the conditions of this study. /e searchstrategy was (head and neck OR oral OR laryngeal ORnasopharyngeal OR pharyngeal OR HNSCC OR LSCC ORNPC OR OSCC) AND (squamous cell carcinoma OR car-cinoma OR tumor OR cancer OR neoplas∗ OR malig-nan∗ ). We filtered the retrieved results according to thefollowing inclusion criteria: (1) the species was Homo sa-piens; (2) samples were from head and neck tissue (includinglaryngeal, oral, tonsil, tongue, pharyngeal, and nasopha-ryngeal tissue), blood, and cell lines; (3) the tumor tissueswere diagnosed as HNSCC; (4) the expression profiling dataof the IL24 mRNA was provided; and (5) the data type wastotal mRNA.

2.3. Statistical Analysis. With SPSS 22.0 software (Chicago,IL, USA), an independent samples t-test was used to in-vestigate the difference in IL24 mRNA expression betweenHNSCC tissues and noncancer tissues based on microarraysand RNA-Seq. Furthermore, the clinical parameters werealso calculated to analyze the factors associated withHNSCC. To visualize these calculations, GraphPad PrismVersion 7.0 (San Diego, CA, USA) was used to create a boxdiagram and receiver operating characteristic (ROC) curvefor each data set. Moreover, the standard mean difference(SMD) and 95% confidence interval (CI) were utilized toshow the different expression levels of the IL24 mRNAbetween HNSCC tissues and noncancer tissues with Stata12.0 software (StataCorp, College Station, TX, USA). /eresults of heterogeneity are reported as a P value or Chi-square test results. A P value less than 0.05 or I2 value greaterthan 50% indicates heterogeneity. /erefore, the randomeffects model was applied. Otherwise, we used the fixedeffects model. In addition, we constructed forest plots to

2 Journal of Oncology

visualize the results. Stata 12.0 was also utilized to constructthe summary ROC (sROC) curves to visualize the capabilityof the IL24 mRNA to distinguish tumor tissues and non-cancer tissues, and the area under the curve (AUC) was usedto evaluate the accuracy of this capability.

3. Results

3.1. 8e Clinical Significance of the IL24 mRNA in OSCC byRNA-Seq. In this study, we downloaded the expressionprofile of the IL24 mRNA in HNSCC from TCGA databaseand extracted the expression data and corresponding clinicalinformation for 341 cases with OSCC samples. Comparedwith the 44 cases with noncancer samples, the OSCCsamples showed significantly higher IL24 mRNA expression(P< 0.0001, Figure 2(m) in the supplementary materials). Inaddition, we analyzed the relevant clinical parameter data,but not found the factors which had a statistically significantcorrelation between the high expression level of the IL24mRNA and OSCC (Table 1). /e Kaplan–Meier curvesuggested that IL24 mRNA expression had no significancewith the prognosis of the patients (Figure 2(p) in the sup-plementary materials).

3.2. 8e Analysis of IL24 mRNA Expression in OSCC byMicroarrays and RNA-Seq Data. In this study, 21 micro-arrays were collected from the GEO database, and RNA-Seqdata were extracted from TCGA database to analyze the IL24mRNA expression between OSCC samples and noncancersamples. /e details of microarrays and RNA-Seq data areshown in Table 2. Compared with noncancer tissues, all thedata sets showed that IL24 was highly expressed in OSCC,and 18 of these data sets had statistical significance(P< 0.05). /e abovementioned results were visualized bybox diagrams and ROC curves (Figures 2(m), 3(a)–3(o), and4(a)–4(f) in the supplementary materials).

To comprehensively analyze the IL24 mRNA ex-pression in OSCC as a whole, all data sets were integratedand analyzed, and the results were combined into a forestplot, which showed that the IL24 mRNA was highlyexpressed in OSCC (SMD � 2.65, 95% CI � 1.66–3.63,P< 0.0001) (Figure 1(a)). /e results showed heteroge-neity, so the random effects model was used (I2 � 96.9%,P< 0.0001; Figure 1(b)). In addition, no significantpublication bias was found (Begg’s test, P � 0.151; Egger’stest, P � 0.599, Figures 1(c) and 1(d)). Furthermore, thesROC curve was used to evaluate the ability of the IL24mRNA to distinguish OSCC from noncancer samples,and the results suggested that the AUC was 0.94(sensitivity � 0.92, specificity � 0.90), which demonstratesdiagnostic ability (Figures 2(a), 2(c), and 2(d)). We alsocalculated the likelihood ratio to synthesize the reliabilityof the sensitivity and specificity. A positive likelihoodratio (+LR) of 9 indicated that the true positive rate ofpatients with OSCC diagnosed with a high expression ofthe IL24 mRNA was 9 times higher than the false positiverate. /e negative likelihood ratio (−LR) was the ratio ofthe false negative rate to the true negative rate. A −LR of

0.09 indicated that the probability of a false negativejudgment was 0.09 times that of a correct negativejudgment (Figure 2(b)).

3.3. 8e Clinical Significance of the IL24 mRNA in LSCC byRNA-Seq. We collected 111 LSCC samples and the relatedclinical information from TCGA database. /e IL24mRNA was significantly higher expressed in LSCC tissuesthan in noncancer tissues (P< 0.0001, Figure 2(n) in thesupplementary materials). We also calculated the clinicalparameters of IL24 mRNA expression, such as age, gender,and pathological grade, in LSCC and noncancer tissues.However, no statistically significant difference was found.Similar to the nonsignificant influence of IL24 on thesurvival of OSCC, no remarkable correlation was notedbetween the IL24 expression and prognosis of LSCC pa-tients (Figure 2(q) in the supplementary materials).

3.4. Analysis of IL24 mRNA Expression in LSCC by Micro-arrays and RNA-Seq Data. To analyze the IL24 mRNA ex-pression in LSCC, we collected six microarrays from the GEOdatabase and RNA-Seq data from TCGA database on LSCC(Table 2)./e calculated results of each data set showed that theIL24 mRNA was highly expressed in LSCC, and six of thesedata sets were statistically significant (P< 0.05, Figures 2(a),2(n), 4(k)–4(o) in the supplementary materials). Due to the

Table 1: /e relationships between IL24 mRNA levels and clini-copathologic features of the 341 cases of OSCC patients detected byRNA-sequencing.

Parameters n Mean ±SD t P valueTissues

OSCC 341 4.055 ±0.2013 10.973

existence of heterogeneity, the random effects model wasused to combine each data set for the overall analysis, andthe forest plot showed that the IL24 mRNA was highlyexpressed in LSCC (SMD � 1.77, 95% CI � 1.10–2.43,P< 0.0001, Figures 3(a) and 3(b)). No publication offsetwas found (Begg’s test, P � 0.133; Egger’s test, P � 0.221,Figures 4(c) and 4(d)). /e sROC curve showed that theIL24 mRNA also had some ability to differentiate LSCCfrom noncancer tissue (AUC � 0.86, sensitivity � 0.81,specificity � 0.82, Figures 4(a), 4(c), and 4(d)). We alsocalculated the likelihood ratio, and the following resultswere obtained: +LR � 5 and −LR � 0.23 (Figure 4(b)).

3.5. Analysis of IL24 mRNA Expression in NPC byMicroarrays. In this study, only four microarrays from theGEO database on the IL24 mRNA expression in NPC werecollected (Table 2), and the relevant clinical parameter datawere not collected. /erefore, only a simple analysis of NPCwas conducted. All four microarrays showed that the IL24mRNA was highly expressed in NPC, but only one wasstatistically significant (GSE53819, P � 0.007, Figures 4(g)–4(j) in the supplementary materials).

Using the fixed effects model (no heterogeneity, I2�11.8%,P � 0.334), the forest plot showed that the IL24 mRNA washighly expressed in NPC (P � 0.039, Figure 5(a)). /e sROC

Table 2: /e basic information of the 39 microarrays and RNA-Seq datasets of IL24 mRNA expression profiling.

ID Subgroup Author CountryHNSCC tissue Non-HNSCC tissue

P valueN Mean ±SD N Mean ±SD

GSE10121 OSCC Sticht C Germany 34 3.027 ±1.612 6 0.404 ±1.194 0.001GSE13601 OSCC Singh B USA 31 5.611 ±1.165 26 4.859 ±0.833 0.007GSE19089 OSCC Smith DI USA 3 6.181 ±0.223 3 5.931 ±0.541 0.501GSE23558 OSCC Ambatipudi S India 27 4.374 ±2.128 5 0.437 ±1.865 0.001GSE25099 OSCC Peng C Taiwan 57 6.368 ±1.275 22 4.073 ±0.395

curve showed the ability of the IL24mRNA to distinguish NPCfrom noncancer (AUC� 0.85, sensitivity� 0.44, specific-ity� 0.86, Figures 5(b)–5(d)). /e following results were alsoobtained: +LR� 3 and −LR� 0.65 (Figure 5(e)).

3.6.8e Clinical Significance of the IL24 mRNA in HNSCC byRNA-Seq. In this study, a total of 502HNSCC samples and thecorresponding clinical data were collected from the TCGAdatabase; these samples included 341OSCC samples, 111 LSCCsamples, and 40 tonsil cancer samples, among which the OSCCand LSCC subgroups were separately analyzed. /e resultsshowed that the IL24 mRNA was highly expressed in HNSCCcompared to noncancer tissue (Figure 2(l) in the supple-mentary materials). Analyzing the clinical parameters, wefound that IL24 mRNA expression was higher in patients withpathological grade 1–2 of HNSCC (Table 3). No relationshipwas observed between IL24 and the survival of HNSCC(Figure 2(o) in the supplementary materials).

3.7. Analysis of IL24 mRNA Expression in HNSCC byMicroarrays and RNA-Seq Data. From the GEO database,we screened and collected a total of 41 microarrays for thisstudy, including 21 microarrays of OSCC, 6 microarrays ofLSCC, 4 microarrays of NPC, and 10microarrays of HNSCCwith an unclassified subgroup. /e RNA-Seq data ofHNSCC (including tonsil, oral cavity, hypopharynx, andlarynx samples) were obtained from the TCGA database(Table 2). /e analysis showed that the IL24 mRNA washighly expressed in all data sets, among which 31 data setshad statistically significant results (Figures 2(a)–2(l), 3(a)–3(o), and 4(a)–4(o) in the supplementary materials). Tocomprehensively analyze IL24 mRNA expression inHNSCC, we combined all the collected data and visualizedthe calculated results with a forest plot. Because of theheterogeneity (I2 � 69.5%, P< 0.0001), the random effectsmodel was used. /e results showed that the IL24 mRNAwas highly expressed in HNSCC compared with noncancertissues (SMD� 1.47, 95% CI� 1.24–1.70, Figure 6(a)), and

StudyID

Begg′s funnel plot with pseudo 95% confidence limits–21.7 0 21.7

4.654.794.274.604.77

4.774.704.584.774.604.714.794.313.944.714.394.524.504.673.764.38

100.00

4.83

SMD (95% CI) %weight

1.68 (0.73, 2.62)0.73 (0.19, 1.27)

0.60 (–1.05, 2.26)1.88 (0.81, 2.95)2.07 (1.48, 2.67)1.58 (1.22, 1.95)2.49 (1.90, 3.08)2.39 (1.58, 3.20)

0.37 (–0.73, 1.47)2.81 (2.19, 3.43)

0.68 (–0.37, 1.73)1.52 (0.72, 2.32)0.95 (0.42, 1.47)3.32 (1.73, 4.92)4.20 (2.04, 6.36)1.69 (0.90, 2.48)1.89 (0.43, 3.34)2.36 (1.14, 3.58)1.91 (0.66, 3.16)1.92 (1.02, 2.83)

2.01 (–0.39, 4.42)20.24 (18.77, 21.70)

2.65 (1.66, 3.63)

GSE10121GSE13601GSE19089GSE23558GSE25099GSE30784GSE31056GSE34106GSE3524GSE37991GSE46802GSE51010GSE55550GSE56532GSE74530GSE9844GSE55549GSE78060GSE75539GSE75538GSE21866TCGA (OSCC)

Overall (I-squared = 96.9%, P = 0.000)NOTE: weights are from randomeffects analysis

(a)

GSE10121GSE13601GSE19089GSE19089GSE23558GSE25099GSE30784GSE31056GSE34106

GSE3524GSE37991GSE46802GSE51010GSE55550GSE56532GSE74530

GSE9844GSE55549GSE78060GSE75539GSE75538GSE21866

TCGA(OSCC)

1.43 1.66 2.65 3.63 3.82

Meta-analysis estimates, given named study is omitted

Lower CI limit

Upper CI limitEstimate

(b)

0 0.5 1 1.5S.e. of: SMD

20

15

10

5

0

SMD

(c)

30

20

10

0

Stan

dard

ized

effec

t

0 2 4 6Precision

Egger′s publication bias plot

(d)

Figure 1: (a) /e forest plot of 22 data sets evaluating the standard mean difference (SMD) of Interleukin 24 (IL24) mRNA expressionbetween oral squamous cell cancer (OSCC) tissues and noncancer tissues based on Gene Expression Omnibus (GEO) and /e CancerGenome Atlas (TCGA) databases. (b) Sensitivity analysis of the expression level of IL24 mRNA in the OSCC and noncancer groups. (c)Begg’s funnel plot for the publication bias test. (d) Egger’s funnel plot for the publication bias test.

Journal of Oncology 5

SROC with confidence and predictive ellipses

1.0

0.5

0.0

Sens

tivity

1.0 0.5 0.0Specificity

Observed data

Summary operating pointSENS = 0.92 [0.83 – 0.97]SPEC = 0.90 [0.85 – 0.93]

SROC curveAUC = 0.94 [0.91 – 0.96]

95% confidence ellipse

95% prediction ellipse

(a)

0.10.20.30.50.7

1

2357

10

20304050607080

909395979899

99.399.599.799.899.9

Pre-

test

prob

abili

ty (%

)

Pre-

test

prob

abili

ty (%

)

10005002001005020105210.50.20.10050.020.010.0050.0020.001

99.999.899.799.599.399

9897959390

80706050403020

10753210.70.50.30.20.1

Likelihood ratio

Fagan′s nomogram

Prior prob (%) = 20LR_Postive = 9Post_Prob_Pos (%) = 69LR_Negative = 0.09Post_Prob_Neg (%) = 2

(b)

Specificity (95% CI)Study ID

TCGA(OSCC)GSE21866GSE75538GSE75539GSE78060GSE55549



Combined

0.73 [0.68 – 0.78]1.00 [0.16 – 1.00]0.86 [0.57 – 0.98]1.00 [0.59 – 1.00]

1.00 [0.63 – 1.00]0.77 [0.56 – 0.91]1.00 [0.54 – 1.00]1.00 [0.69 – 1.00]0.63 [0.54 – 0.71]0.96 [0.86 – 0.99]0.43 [0.10 – 0.82]0.90 [0.76 – 0.97]0.31 [0.11 – 0.59]1.00 [0.88 – 1.00]1.00 [0.85 – 1.00]0.81 [0.75 – 0.87]0.95 [0.85 – 0.99]0.96 [0.81 – 1.00]1.00 [0.29 – 1.00]0.61 [0.42 – 0.78]1.00 [0.90 – 1.00]

0.92 [0.83 – 0.97]

0.92 [0.75 – 0.99]

I2 = 90.47 [87.46 – 93.48]

Q = 220.41, df = 21.00, P = 0.00

0.1 1.0Sensitivity

(c)

Study ID Specificity (95% CI)

TCGA(OSCC)GSE21866GSE75538GSE75539GSE78060GSE55549



Combined

0.91 [0.78 – 0.97]1.00 [0.29 – 1.00]0.93 [0.66 – 1.00]0.88 [0.47 – 1.00]

1.00 [0.40 – 1.00]1.00 [0.74 – 1.00]1.00 [0.54 – 1.00]1.00 [0.54 – 1.00]0.94 [0.70 – 1.00]0.88 [0.47 – 1.00]0.88 [0.47 – 1.00]0.93 [0.80 – 0.98]1.00 [0.40 – 1.00]0.73 [0.45 – 0.92]0.79 [0.68 – 0.88]0.98 [0.88 – 1.00]0.91 [0.71 – 0.99]0.80 [0.28 – 0.99]0.67 [0.09 – 0.99]0.85 [0.65 – 0.96]0.83 [0.36 – 1.00]

0.90 [0.85 – 0.93]

1.00 [0.40 – 1.00]

I2 = 45.92 [18.88 – 72.96]

Q = 38.83, df = 21.00, P = 0.01

0.1 1.0Sensitivity

(d)

Figure 2: (a)/e summary receiver operating characteristic curves analysis of IL24mRNA for discriminating OSCC from noncancer tissuesbased on GEO and TCGA databases. (b) Prior probability and postprobability positive and negative of the included studies. /e priorprobability, postprobability positive, and postprobability negative reached 20%, 69%, and 2%, respectively (LR: likelihood ratio). (c, d)Sensitivity and specificity values of the included studies. /e sensitivity and specificity values of the included studies were 0.92 (95%CI� 0.83−0.97) and 0.90 (95%CI� 0.85−0.93), respectively.


there was no publication bias (Begg’s test, P � 0.176; Egger’stest, P � 0.201; Figures 6(b) and 6(c)). In addition, weconstructed an sROC curve, whose AUC demonstrated theability of IL24 mRNA expression to distinguish HNSCCfrom noncancer tissue (AUC� 0.93, 95% CI� 0.91−0.95,Figure 7(a)). /e sensitivity and specificity were 0.83, 95%CI� 0.76−0.89 and 0.89, 95% CI� 0.85−0.92, respectively(Figures 7(c) and 7(d)). /e following results were alsoobtained: +LR� 8 and −LR� 0.19 (Figure 7(b)).

4. Discussion

IL24 mRNA is known to have an apoptotic effect oncancer cells but not on noncancer cells [22, 23]. IL24mRNA could induce autophagy as a protective mecha-nism, while 3-methyladenine (3-MA) inhibition ofautophagy significantly enhanced the anticancer effect ofIL24 in OSCC [18]. /ere also was a study that showedthat the transfection of miR-205 into human oral car-cinoma (KB) cells strongly induced the IL24 mRNA,which targeted the IL24 promoter directly to induce geneexpression and had significant therapeutic potential toturn on silenced tumor suppressor genes [24]. Adeno-virus-mediated IL24 mRNA therapy could enhance the invivo and in vitro antitumor effects of radiotherapy forNPC [20, 21]. A study of Bifidobacterium as a carrier ofthe IL24 mRNA to treat HNSCC in mice showed that therecombinant strain Bifidobacterium breve-IL24 had a

stronger ability to inhibit tumor growth and induce tu-mor cell apoptosis [19]. Although there is no suitable wayto conduct clinical trials of the IL24 mRNA as a targetedtherapy for HNSCC and its subgroups, the current studiessuggest that the IL24 mRNA could be a potential treat-ment. /e body will be protected by regulating the ex-pression of the IL24 mRNA as an immune factor duringcancer. Many studies have found that IL24 can beexpressed through a variety of immune cells—especiallyT-helper cells (/)2 cells and tissue cells, includingkeratinocytes, epithelial cells, and endothelial cells, andplays an important role in host defense, tissue protection,and autoimmune response regulation [25, 26]. It has alsobeen studied in many cancers, such as prostate cancer,kidney cancer, osteosarcoma, melanoma, and coloncancer [13, 14, 16]. Induction of endoplasmic reticulum (ER)stress is one of the main mechanisms by which IL24 mRNAinduces cell death. Compared with normal cells, cancer cellshave higher levels of ER stress, which makes them moresusceptible to ER stress-mediated cell death triggered by IL24mRNA. Another possible explanation is that IL24 mRNAmediates different cell killing effects between normal andcancer cells depending on reactive oxygen species (ROS), and anumber of studies have shown that cancer cells have higherlevels of basic ROS than normal cells. /erefore, comparedwith normal cells, drugs such as IL24mRNA that enhance ROSproduction can overcome the natural antioxidant effect incancer cells more effectively, leading to cell death [9, 27].However, neither the correlation between IL24 mRNA ex-pression and HNSCC has been analyzed in the literature norhas there been a detailed study of IL24 mRNA expression inHNSCC and its subgroups or a cross-sectional comparisonbetween them. /erefore, this study mainly analyzed IL24mRNA expression and the corresponding clinical significancein HNSCC and its subgroups at the mRNA level.

HNSCC is a squamous cell tumor composed of varioussubgroups, among which the most common subgroupsare OSCC, NPC, and LSCC [28, 29]. /erefore, we mainlyanalyzed these three common subgroups and HNSCC as awhole group in this study. To more accurately illustratethe clinical significance of the IL24 mRNA in HNSCC andits subgroups, substantial IL24 mRNA expression datawere selected by searching several databases. We ulti-mately collected a total of 41 microarrays and RNA-seq onHNSCC from the GEO and TCGA databases. Amongthem, 21 were for OSCC; 4 were for NPC; and 6 were forLSCC; the others were for an unclassified subgroup ofHNSCC. A total of 1,564 HNSCC tissue samples (in-cluding 1060 cases of OSCC, 86 cases of NPC, and 181cases of LSCC) and 603 noncancer tissue samples, as wellas the corresponding clinical information regarding 502HNSCC patients (including 341 cases of OSCC and 111cases of LSCC), were included. /e inclusion of manysamples made the results of this analysis more reliable andmore detailed. We utilized the microarrays and RNA-seqdata to analyze the difference in IL24 mRNA expression ineach subgroup and HNSCC compared with noncancertissue. /en, we investigated the relationship betweenIL24 mRNA expression and clinical parameters. /e

Table 3: /e relationships between IL24 mRNA levels and clini-copathologic features of the 502 cases of HNSCC patients detectedby RNA-sequencing.

Parameters n Mean ±SD t P valueTissueHNSCC 502 4.035 ±0.1973 10.504

results showed that the IL24 mRNA was highly expressedin HNSCC and its subgroups and had the ability to dis-tinguish between cancer and noncancer tissues. InHNSCC and its subgroups, the results showed that thesepatients who highly expressed the IL24 mRNA seemed tohave a better prognosis.

In this study, the IL24 mRNA had significantly higherexpression in OSCC, as well as in LSCC, compared tononcancer tissues, and both comparisons had significantstatistical differences (P< 0.0001). /e sROC curve showedthe AUC of OSCC was 0.94, while the AUC of LSCC was0.86, which both suggested the IL24 mRNA had the ability todistinguish between tumor tissues and noncancer tissues. Inaddition, we also studied the relevant clinical parameters ofIL24 mRNA expression in OSCC and LSCC, such as age,gender, and pathological stage, but did not find statisticallysignificant indicators. /en, we conducted a subgroupanalysis of NPC. Compared with other types of HSNCC, thesusceptibility factors leading to NPC somewhat differed. Acommon factor is Epstein–Barr virus (EBV) infection, and arare factor is HPV infection; although some studies havereported cases, there is no significant correlation betweenHPV infection and NPC [30–32]. We found that IL24 washighly expressed in NPC (P � 0.039). /ese findings suggestthat NPC, like other HNSCC tissues, may be related to highexpression of the IL24 mRNA, which is worth further studyof the molecular mechanism in the future.

We combined all the collected data to analyze IL24mRNA expression in HNSCC as a whole. IL24 was highlyexpressed in HNSCC and had the ability to distinguishHNSCC from noncancer tissue. Analyzing the clinicalparameters, we found that IL24 mRNA expression washigher in patients with pathological grade 1–2 ofHNSCC. IL24 was reported to be a potential prognosticbiomarker and an indicator of malignancy in HNSCC in2014 [33]. In vitro tests showed that high expression ofmda-7/IL24 can upregulate the expression of the epi-thelial terminal differentiation markers cytokeratin(KRT) 1, KRT4, KRT13, and phosphorylated endoplas-mic reticulum stress protein (p)-EIF2a. However, it alsomay be part of the underlying mechanism involved inmda-7-mediated HNSCC differentiation and apoptosisvia downregulation of the epithelial proliferative markersKRT5, KRT14, integrin β4, and antiapoptosis proteinBcl-2. Although this study analyzed the potential path-way mechanism of high expression of the IL24 mRNA inHNSCC as a whole part, no other study has conductedthe same analysis for the IL24 mRNA in a subgroup ofHNSCC. /erefore, we mainly analyzed the expression ofthe IL24 mRNA in OSCC, LSCC, and NPC, which was thesame as that in HNSCC, and they all demonstrated highIL24 mRNA expression. /is finding also suggested thatthe expression mechanism of the IL24 mRNA in eachsubgroup may be the same as that in HNSCC, which is

StudyID SMD (95% CI)

%weight

GSE51985GSE59102GSE29330GSE84957

GSE117005GSE143224TCGA (LSCC)Overall (I-squared = 72.0%, P = 0.002)

NOTE: Weights are from random effects analysis

15.66

13.6817.18

14.30

6.5511.6420.98

0.88 (–0.04, 1.80)1.91 (1.13, 2.68)1.27 (0.15, 2.39)1.49 (0.43, 2.54)

3.67 (1.48, 5.85)3.79 (2.44, 5.14)

100.001.11 (0.74, 1.48)1.77 (1.10, 2.43)

–5.85 0 5.85

(a)

GSE51985

GSE59102

GSE29330

GSE84957

TCGA (LSCC)

GSC117005

GSC143224

Meta-analysis random-effects estimates (linear from) Study ommited

0.94 1.10 1.77 2.43 2.80

(b)

–2

0

2

4

SMD

0 0.5 1S .e. of: SMD

Begg’s funnel plot with pseudo 95% confidence limits

(c)

Egger’s publication bias plot6

2

0

4

–2

Stan

dard

ized

effec

t

0 2 4 6Precision

(d)

Figure 3: (a) /e forest plot of 7 data sets. (b) Sensitivity analysis of analysis of the expression level of IL24 mRNA in the LSCC andnoncancer groups. (c) Begg’s funnel plot for the publication bias test. (d) Egger’s funnel plot for the publication bias test.


1.0

0.5

0.0

Sens

itivi

ty

1.0 0.5 0.0Specificity


Observed dataSummary operating pointSENS = 0.81 [0.70 – 0.89]SPEC = 0.82 [0.72 – 0.89]SROC curveAUC = 0.86 [0.83 – 0.89]

95% confidence ellipse95% prediction ellipse

(a)

Likelihood ratio

0.10.20.30.50.7

1

0.10.20.30.50.71

2357

10

235710

20304050607080 20

304050607080

9093959798

9093959798

9999.399.599.799.899.9

9999.399.599.799.899.9

Pre-

test

prob

abili

ty (%

)

Pre-

test

prob

abili

ty (%

)

10005002001005020102510.50.20.10.050.020.010.0050.0020.001

Fagan′s nomogram

Prior prob (%) = 20LR_Postive = 5Post_Prob_Pos (%) = 53LR_Negative = 0.23Post_Prob_Neg (%) = 5

(b)

Study ID

TCGA (LSCC)

GSE143224

GSE117005

GSE84957

GSE29330

GSE59102

GSE51985

Combined

Sensitivity (95% CI)

0.72 [0.63 – 0.80]

0.92 [0.62 – 1.00]

0.80 [0.28 – 0.99]

0.75 [0.40 – 0.97]

0.77 [0.46 – 0.95]

0.93 [0.77 – 0.99]

0.80 [0.44 – 0.97]

0.81 [0.70 – 0.89]

I2 = 28.93 [0.00 – 88.96]

Q = 8.44, df = 6.00, P = 0.21

0.3 1.0Sensitivity

(c)

Study ID

TCGA (LSCC)

GSE143224

GSE117005

GSE84957

GSE29330

GSE59102

GSE51985

Combined

Sensitivity (95% CI)

0.82 [0.67 – 0.92]

0.77 [0.46 – 095]

0.80 [0.28 – 0.99]

0.89 [0.52 – 100]

1.00 [0.48 – 1.00]

0.92 [0.64 – 1.00]

0.60 [0.26 – 0.88]

0.82 [0.72 – 0.89]

I2 = 5.38 [0.00 – 100.00]

Q = 6.34, df = 6.00, P = 0.39

0.3 1.0Sensitivity

(d)

Figure 4: (a)/e summary receiver operating characteristic curves analysis of IL24 mRNA for discriminating LSCC from noncancer tissuesbased on GEO and TCGA databases. (b) Prior probability and postprobability positive and negative of the included studies. /e priorprobability, postprobability positive, and postprobability negative reached 20%, 53%, and 5%, respectively (LR: likelihood ratio). (c, d) 0.81(95% CI� 0.70−0.89) and 0.82 (95% CI� 0.72−0.89), respectively.



%weight

GSE12452

GSE13597

GSE53819

0.07 (–0.65, 0.78) 35.56

12.53

37.83

14.08

100.00

0.33 (–0.87, 1.53)

0.95 (0.26, 1.64)

0.16 (–0.97, 1.30)

0.45 (0.02, 0.87)

GSE64634

Overall (I–square = 11.8%, P = 0.334)

–1.64 1.640

(a)

1.0

0.5

0.01.0 0.5 0.0

Sens

itivi

ty


Specificity



(b)

GSE12452

Combined

0.1 0.8Sensitivity

GSE13597

GSE53819

GSE64634

Study ID Sensitivity (95% CI)

0.58 [0.28 – 0.85]

0.61 [0.36 – 0.83]

0.28 [0.12 – 0.49]

0.44 [0.31 – 0.58]

Q = 6.08, df = 3.00, p = 0.11

I2 = 50.65 [0.00 – 100.00]

0.39 [0.22 – 0.58]

(c)

GSE12452

Combined

0.1 0.8Sensitivity

GSE13597

GSE53819

GSE64634

Study id Specificity (95% CI)

0.75 [0.19 – 0.99]

0.89 [0.65 – 0.99]

1.00 [0.29 – 1.00]

0.86 [0.69 – 0.94]

Q = 1.27, df = 3.00, P = 0.74

I2 = 0.00 [0.00 – 100.00]

0.80 [0.44 – 0.97]

(d)

Figure 5: Continued.


Fagan’s nomogram0.10.20.30.50.7

1

2357

10

20304050607080

9093959798

9999.399.599.799.8

99.9 0.1

0.20.30.50.71

235710

20304050607080

9093959798

9999.399.599.799.899.9

Pre-

test

prob

abili

ty (%

)

Post-

test

prob

abili

ty (%

)

Likelihood ratio

10005002001005020105210.50.20.10.050.020.010.0050.0020.001

Prior prob (%) = 20LR_Positive = 3Post_Prob_Pos (%) = 43LR_Negative = 0.65Post_Prob_Peg (%) = 14

(e)

Figure 5: (a) /e forest plot of 4 data sets evaluating the standard mean difference (SMD) of Interleukin 24 (IL24) mRNA expressionbetween nasopharyngeal carcinoma (NPC) tissues and noncancer tissues based on Gene Expression Omnibus (GEO). (b) /e summaryreceiver operating characteristic curves analysis of IL24 mRNA for discriminating NPC from noncancer tissues based on GEO database. (c,d) Sensitivity and specificity values of the included studies. /e sensitivity and specificity values of the included studies were 0.44 (95%CI� 0.31−0.58) and 0.86 (95% CI� 0.69−0.94), respectively. (e) Prior probability and postprobability positive and negative of the includedstudies. /e prior probability, postprobability positive, and postprobability negative reached 20%, 43%, and 14%, respectively (LR:likelihood ratio).


%weight

GSE10121GSE13601GSE19089GSE23558GSE25099GSE30784GSE31056GSE34106GSE3524GSE37991GSE46802GSE51010GSE55550GSE56532GSE74530GSE9844GSE55549GSE78060GSE75539GSE75538GSE21866GSE12452GSE13597GSE53819GSE64634GSE51985GSE59102GSE29330GSE84957

GSE10774

GSE117005GSE143224

GSE13397GSE13398GSE33205GSE55546GSE58911

GSE83519GSE107591GSE73330TCGA

–6.36 0 6.36

1.68 (0.73, 2.62) 2.390.73 (0.19, 1.27) 3.28

0.60 (–1.05, 2.26) 1.31

0.37 (–0.73, 1.47) 2.09

0.68 (–0.37, 1.73) 2.201.52 (0.72, 2.32) 2.700.95 (0.42, 1.47) 3.303.32 (1.73, 4.92) 1.38

GSE6631 0.76 (0.15, 1.38) 3.12

4.20 (2.04, 6.36) 0.891.69 (0.90, 2.48) 2.731.89 (0.43, 3.34) 1.542.36 (1.14, 3.58) 1.891.91 (0.66, 3.16) 1.841.92 (1.02, 2.83) 2.47

2.01 (–0.39, 4.42) 0.750.07 (–0.65, 0.78) 2.890.33 (–0.87, 1.53) 1.920.95 (0.26, 1.64) 2.94

0.16 (–0.97, 1.30) 2.040.88 (–0.04, 1.80) 2.441.91 (1.13, 1.68) 2.751.27 (0.15, 2.39) 2.061.49 (0.43, 2.54) 2.183.67 (1.48, 4.85) 0.883.97 (2.44, 5.14) 1.69

1.23 (–0.03, 2.50) 1.83

0.99 (–0.06, 2.04) 2.201.27 (0.18, 2.36) 2.12

0.82 (0.31, 1.33) 3.341.30 (0.07, 2.53) 1.881.59 (0.77, 2.42) 2.64

1.12 (0.48, 1.75) 3.061.58 (0.92, 2.24) 3.020.85 (0.01, 1.69) 2.621.17 (0.86, 1.49) 3.711.47 (1.24, 1.70) 100.00

2.81 (2.19, 3.43) 3.10

1.88 (0.81, 2.95) 2.162.07 (1.48, 2.67) 3.171.58 (1.22, 1.95) 3.632.49 (1.90, 3.08) 3.172.39 (1.58, 3.20) 2.67

Overall (I-squared = 69.5%, P = 0.000)NOTE: weights are from random effects analysis

(a)

4

Begg’s funnel plot with pseudo 95% confidence limits

2

0

SMD

–2

0 0.5S.e.of; SMD

1 1.5

(b)



Egger’s publication bias plot

Stan

dard

ized

effec

t

10

5

0

0 2Precision

4 6

(c)

Figure 6: (a)/e forest plot of 42 data sets. (b) Begg’s funnel plot for the publication bias test. (c) Egger’s funnel plot for the publication biastest.

1.0

0.5

0.01.0 0.5 0.0

Sens

itivi

ty


Specificity



(a)

Fagan’s nomogram0.10.20.30.50.7

1

2357

10

20304050607080

909395979899

99.399.599.799.899.9 0.1

0.20.30.50.71235710

20304050607080

9093959798

9999.399.599.799.899.9

Pre-

test

prob

abili

ty (%

)

Post-

test

prob

abili

ty (%

)

Likelihood ratio

10005002001005020105210.50.20.10.050.020.010.0050.0020.001

Prior prob (%) = 20LR_Positive = 8Post_Prob_Pos (%) = 66LR_Negative = 0.19Post_Prob_Neg (%) = 4

(b)



worth a detailed study in the future to provide moreaccurate targeted therapy candidates for each subgroup ofHNSCC. In addition, we analyzed the Kaplan–Meiercurves in HNSCC and each subgroup, all of which sug-gested that IL24 mRNA expression had no significantcorrelations with the status of prognosis, which needs tobe verified with more cohorts. IL24 can protect the bodyvia the induction of tumor cell apoptosis and inhibition ofangiogenesis [10, 11] in cancers such as colorectal ade-nocarcinoma and hepatic carcinoma [34, 35]. IL24 hasalso been found in many diseases involving inflammation,such as cardiovascular disease, rheumatoid arthritis, tu-berculosis, and viral infections, and may play a role inpsoriasis [15, 36]. In addition, many studies have shownthat IL24 mRNA can be used as a potential tumortreatment [37, 38]. All these characteristics suggest thatthe IL24 mRNA may provide a potential idea for thetreatment of HNSCC and its subgroups, which is sig-nificant for further research with a larger sample size.

However, this study also had some limitations. /erewas a large difference in the number of samples betweencancer and noncancer. Although appropriate statistical

methods were used for analysis, the error of calculationresults cannot be completely avoided. We did not furtherstudy the molecular mechanism and pathway of IL24mRNA expression in each subgroup, which is worthfurther study in the future. /e expression level of theIL24 mRNA in the bodily fluid of patients with HNSCCrepresents its potential for early diagnosis in clinicalsettings, which also requires further testing.

In conclusion, the IL24 mRNA was highly expressedin HNSCC, as well as in its subgroups—OSCC, LSCC,and NPC. According to the results of sROC, IL24 mRNAalso had the ability to distinguish between cancer andnoncancer tissues in HNSCC and its subtypes, respec-tively. /e IL24 mRNA level may be used as a potentialmarker for HNSCC screening and was worthy of furtherverification in the future. We hope that analysis of theIL24 mRNA expression pathway and molecular mech-anism will be conducted and explored the relationship ofthe IL24 mRNA with the development of HNSCC anddisease progression with a larger sample size. In addition,we hope to provide a new idea for the treatment of theIL24 mRNA in HNSCC and its subgroups.

0.1 1.0Sensitivity

GSE73330GSE107591

GSE83519GSE6631


GSE84957GSE117005GSE143224

GSE29330GSE59102GSE51985GSE64634GSE53819GSE13597GSE12452GSE21866GSE75538GSE75539GSE78060GSE55549



Combined

TCGA

Study ID Sensitivity (95% CI)

0.69 [0.64 – 0.73]0.58 [0.28 – 0.85]0.58 [0.37 – 0.78]0.68 [0.45 – 0.86]

0.93 [0.68 – 1.00]0.77 [0.55 – 0.92]

0.83 [0.52 – 0.98]0.55 [0.39 – 0.70]0.75 [0.35 – 0.97]0.88 [0.47 – 1.00]0.70 [0.35 – 0.93]

0.78 [0.40 – 0.97]0.77 [0.46 – 0.95]

0.92 [0.62 – 1.00]0.80 [0.28 – 0.99]

0.93 [0.77 – 0.99]0.80 [0.44 – 0.97]0.58 [0.28 – 0.85]0.61 [0.36 – 0.83]0.28 [0.12 – 0.49]0.39 [0.22 – 0.58]1.00 [0.16 – 1.00]0.86 [0.57 – 0.98]1.00 [0.59 – 1.00]0.92 [0.75 – 0.99]1.00 [0.63 – 1.00]0.77 [0.56 – 0.91]1.00 [0.54 – 1.00]1.00 [0.69 – 1.00]0.63 [0.54 – 0.71]0.96 [0.86 – 0.99]0.43 [0.10 – 0.82]0.90 [0.76 – 0.97]0.31 [0.11 – 0.59]1.00 [0.88 – 1.00]1.00 [0.85 – 1.00]0.81 [0.75 – 0.87]0.95 [0.85 – 0.99]0.96 [0.81 – 1.00]1.00 [0.29 – 1.00]0.61 [0.42 – 0.78]1.00 [0.90 – 1.00]

0.83 [0.76 – 0.89]Q = 266.18, df = 41.00, P = 0.00I2 = 84.60 [80.58 – 88.61]

(c)

Specificity (95% CI)

0.91 [0.78 – 0.97]0.92 [0.62 – 1.00]1.00 [0.85 – 1.00]0.82 [0.60 – 0.95]

0.93 [0.68 – 1.00]1.00 [0.40 – 1.00]0.96 [0.80 – 1.00]0.88 [0.47 – 1.00]0.88 [0.47 – 1.00]1.00 [0.40 – 1.00]

0.80 [0.28 – 0.99]0.77 [0.46 – 0.95]

0.89 [0.52 – 1.00]1.00 [0.48 – 1.00]0.92 [0.64 – 1.00]0.60 [0.26 – 0.88]0.75 [0.19 – 0.99]0.89 [0.65 – 0.99]1.00 [0.29 – 1.00]0.80 [0.44 – 0.97]1.00 [0.29 – 1.00]0.93 [0.66 – 1.00]0.88 [0.47 – 1.00]1.00 [0.40 – 1.00]1.00 [0.40 – 1.00]1.00 [0.74 – 1.00]1.00 [0.54 – 1.00]

0.55 [0.32 – 0.76]

1.00 [0.54 – 1.00]0.94 [0.70 – 1.00]0.88 [0.47 – 1.00]0.88 [0.47 – 1.00]0.93 [0.80 – 0.98]1.00 [0.40 – 1.00]0.73 [0.45 – 0.92]0.79 [0.68 – 0.88]0.98 [0.88 – 1.00]0.91 [0.71 – 0.99]0.80 [0.28 – 0.99]0.67 [0.09 – 0.99]0.85 [0.65 – 0.96]0.83 [0.36 – 1.00]

0.89 [0.85 – 0.92]Q = 79.39, df = 41.00, P = 0.00I2 = 48.36 [29.79 – 66.93]

0.1 1.0Specificity

GSE73330GSE107591

GSE83519GSE6631


GSE84957GSE117005GSE143224

GSE29330GSE59102GSE51985GSE64634GSE53819GSE13597GSE12452GSE21866GSE75538GSE75539GSE78060GSE55549



Combined

TCGA

Study ID

(d)

Figure 7: (a) /e summary receiver operating characteristic curves analysis of IL24 mRNA for discriminating HNSCC from noncancertissues based on GEO and TCGA databases. (b) Prior probability and postprobability positive and negative of the included studies./e priorprobability, postprobability positive, and postprobability negative reached 20%, 66%, and 4%, respectively (LR: likelihood ratio). (c) and(d) Sensitivity and specificity values of the included studies. /e sensitivity and specifcity were 0.83, 95%CI� 0.76–0.89 and 0.89,95%CI� 0.85–0.92, respectively.


Data Availability

Data used for analysis were extracted from the publiclyavailable databases—the Gene Expression Omnibus (GEO)and the Cancer Genome Atlas (TCGA) databases. /e in-formation of microarrays and RNA-Seqs data used tosupport the findings of this study are included within article.Please see Table 2 for details.

Conflicts of Interest

/e authors declare that they have no conflicts of interest.

Authors’ Contributions

Lan-Lan Qiu and Xiao-Guohui Zhang contributed equally asthe co-first authors.

Acknowledgments

/is study was supported by the National Natural ScienceFoundation of China (31760319), the Guangxi First-classDiscipline Project for Basic medicine Sciences (No.GXFCDP-BMS-2018), Guangxi Degree and PostgraduateEducation Reform and Development Research Projects,China (JGY2019050), 2019 Guangxi Medical UniversityEducation and Teaching Reform Project (2019XJGZ04), andGuangxi Zhuang Autonomous Region Health and FamilyPlanning Commission Self-financed Scientific ResearchProject (Z20180979).

Supplementary Materials

Figure 1: flow chart of the study design. Supplementary ma-terials: Figure 2: (a) the expression data of IL24 mRNA and thecorresponding ROC curves in LSCC tissues fromGEOdatabase.(b–k) /e expression data of IL24 mRNA and the corre-sponding ROC curves in unclassified subgroups HNSCC tissuesin 10microarrays fromGEOdatabase. (l–n)/e expression dataof IL24 mRNA and the corresponding ROC curves in HNSCCand its subgroups (OSCC and LSCC) tissues in RNA-Seq byTCGA database. (o) /e Kaplan–Meier survival curve of IL24mRNA in the HNSCC by RNA-Seq. (p) /e Kaplan–Meiersurvival curve of IL24mRNA in theOSCCby RNA-Seq. (q)/eKaplan–Meier survival curve of IL24 mRNA in the LSCC byRNA-Seq.Note: receiver operating characteristic (ROC); headand neck squamous cell carcinoma (HNSCC); oral squamouscell carcinoma (OSCC); laryngeal squamous cell carcinoma(LSCC). Figure 3: (a–o) the expression data of IL24 mRNA andthe corresponding ROC curves in OSCC tissues in 15 micro-arrays from the GEO database, Figure 4: (a–f) the expressiondata of IL24 mRNA and the corresponding ROC curves inOSCC tissues in 6 microarrays from GEO database. (g–j) /eexpression data of IL24 mRNA and the corresponding ROCcurves in NPC tissues in 4 microarrays from GEO database.(k–o) /e expression data of IL24 mRNA and the corre-sponding ROC curves in LSCC tissues in 5 microarrays fromGEO database. (Supplementary Materials)

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