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Inhibition of PDCD6 Induces Cell Proliferation and Reduces Apoptosis in Human Epithelial Ovarian Cancer Cells
Yan Huang, Xiaohua Wu
Department of Gynecology, Cancer Center, Fudan University
Yan Huang ( 310108197903152242)
Correspondence: Xiaohua Wu
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Ovarian epithelial cancer is the leading killer among all gynecological malignancies.
Recent findings suggested that apoptosis and apoptosis-linked genes may play an important role in the development and progression of ovarian cancer (la Cour JM, et al. Mol Oncol. 2008).
Programmed cell death gene 6 (PDCD6), also known as apoptosis-linked gene-2 (ALG-2), is a 22 kDa penta-EF-hand type Ca2+-binding protein that was first reported by Vito in 1996. It is well recognized as an apoptotic mediator because PDCD6 expression is required for apoptosis following numerous death stimuli (Vito P, et al. Science. 1996).
Backgrounds
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MethodsPoliferation
BrdU
Flow cytometry
RT-PCR, Western blot
Design of siRNA and construction of recombi-nant plasmids
Flow cytometry
Poliferation
Apoptosis
PDCD6
RT-PCR, Western blot
RT-PCR, Western blot
RT-PCR, Western blot
BrdU, Flow cytometry
Effects of cisplatin on ovarian cancer cells SKOV-3 and OVCAR-3
Transfection
SKOV-3 and OVCAR-3
Cell cycle
Apoptosis
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Results RNAi inhibited PDCD6 expression in ovarian
cancer cells
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Inhibition of PDCD6 prohibited apoptosis and increased cell proliferation
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Inhibition of PDCD6 induced cell cycle
Cell cycle phase
Transfection group G0/G1(%) S(%) G2/M(%)
SKOV-3 control 75.23±1.17 16.87±0.58 7.90±1.39
SKOV-3 pRT-neg 76.73±1.53 15.40±1.21 7.87±1.45
SKOV-3 pRT-pdcd 56.77±1.48a 26.27±1.11a 16.97±2.56a
OVCAR-3 control 79.90±1.35 17.17±1.11 2.93±1.08
OVCAR-3 pRT-neg 78.27±1.00 17.30±1.13 4.43±2.05
OVCAR-3 pRT-pdcd 59.27±0.90b 22.17±1.80b 18.57±1.53b
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• Cisplatin inhibited proliferation and increased apoptosis
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• PDCD6 RNAi partly blocked the apoptotic effect and the anti-proliferative effect induced by cisplatin
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Conclusions• 1. Inhibition of PDCD6 by RNAi induced cell
proliferation and cell cycle in SKOV-3 and OVCAR-3 cells by up-regulating cyclin D1, cyclin B1, cyclin E, and proliferating cell nuclear antigen (PCNA).
• 2. Inhibition of PDCD6 by RNAi reduced apoptosis in SKOV-3 and OVCAR-3 cells by decreasing cleaved caspase-3, cleaved caspase-7, cleaved caspase-8, Bax, death receptor 5 (DR5 ) and death receptor 4(DR4) and increasing Bcl-2.
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• 3. Cisplatin increased cell apoptosis and prohibited cell proliferation while the mRNA and protein expression of PDCD6 was up-regulated.
• 4. PDCD6 RNAi partly blocked the apoptotic effect and the anti-proliferative effect induced by cisplatin. Cisplatin may inhibit proliferation and increase apoptosis partly through PDCD6.
• 5. PDCD6 plays an important role in the promotion and progression of ovarian cancer. PDCD6 is a candidate as a new target for ovarian cancer treatment.
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