Survivin反义寡核苷酸对人卵巢癌细胞株IL-6/STAT3信号通路的影响和意义
Effects and values of antisurvivin oligonucleotides on IL-6/STAT3 signaling pathway of ovarian cancer cell line

摘要：目的 通过分析人卵巢癌细胞株转染Survivin反义寡核苷酸(ASODN)后IL-6/STAT3信号通路及其下游相关癌基因的表达以及其侵袭能力的改变，探讨Survivin ASODN在抑制人卵巢癌细胞转移中的可能作用、机制和临床价值。方法 脂质体(LipofectamineTM2000)介导Survivin ASODN转染人卵巢癌SKOV3细胞株(实验组)，同时设空脂质体对照，Transwell小室检测细胞株侵袭能力的改变，荧光定量PCR检测两组细胞株中白细胞介素6(IL-6)、信号转导与活化因子3(STAT3)、Survivin、血管内皮生长因子(VEGF)基因的表达，Western blot检测IL-6、STAT3、磷酸化的信号转导与活化因子3(p-STAT3)、Survivin、VEGF-A蛋白的表达。结果 实验组细胞株中IL-6、STAT3、Survivin、VEGF基因表达均较对照组明显下调(P<0.05)；实验组细胞株中IL-6、STAT3、p-STAT3、Survivin、VEGF-A蛋白表达均较对照组明显下调(P<0.05)。实验组细胞株侵袭能力较对照组明显下降(P<0.01)。结论 ASODN靶向抑制Survivin可有效降低人卵巢癌细胞侵袭转移能力，抑制IL-6/STAT3信号通路及其下游相关致癌基因的表达活性。Survivin ASODN可能在防治卵巢癌复发和转移治疗中有临床价值。
ABSTRACT: Objective To analyze the effects of antisurvivin oligonucleotide (Survivin ASODN) on IL-6/STAT3 signaling pathway and down-stream cancer genes of ovarian cancer SKOV3 cell line and to detect the changes of invasive ability of cell line in order to explore the role of Survivin ASODN in reducing metastasis and recurrence of ovarian cancer and its potential clinical value. Methods ASODN was transfected into SKOV3 cells by lipofectamineTM 2000 in ASODN group, and lipofectamineTM 2000 was introduced in control group. The invasive ability in ASODN and control groups was detected by Transwell chamber. The expressions of interieukin-6(IL-6), signal transducer and activator of transcription 3(STAT3), Survivin, and vascular endothelial growth factor (VEGF) in ovarian cancer cell line at mRNA level were detected by Real-time PCR. The expressions of IL-6, STAT3, signal transducer and activator of transcription 3 phosphorylation (p-STAT3), Survivin, and VEGF-A in ovarian cancer cell line at protein level were detected by Western blot. Results The expressions of IL-6, STAT3, Survivin, and VEGF in ovarian cancer cell line at mRNA level were significantly down-regulated in ASODN group (P<0.05). IL-6, STAT3, p-STAT3, Survivin and VEGF-A protein levels in ovarian cancer cell line were significantly down-regulated in ASODN group (P<0.05). The invasive ability was significantly reduced by ASODN (P<0.01). Conclusion ASODN targeting Survivin could reduce the invasive ability of ovarian cancer cell line. The mechanism may be blocking IL-6/STAT3 signaling pathway and its down-stream cancer genes of ovarian cancer cell line. ASODN targeting Survivin may have clinical value in preventing and treating the metastasis and recurrence of ovarian cancer