Survivin反义寡核苷酸对卵巢癌细胞株凋亡和侵袭、迁移的影响

目的:观察survivin反义寡核苷酸(ASODN)对人卵巢癌细胞株SKOV3凋亡和侵袭、迁移的影响,探讨其作用的分子机制和临床价值。方法:用脂质体LipofectamineTM2000介导ASODN转染人卵巢癌细胞株SKOV3,Transwell小室检测细胞侵袭和迁移能力的改变,流式细胞学技术检测细胞凋亡指数及survivin、Smac、VEGF蛋白表达改变,逆转录酶链反应(RT-PCR)检测survivin、Smac/DIABLO、VEGF基因表达改变。结果:脂质体LipofectamineTM2000介导ASODN转染人卵巢癌细胞株SKOV3,FCM及共聚焦显微镜(SLM510)检测结果显示转染率>95%。用600 nmol/L ASODN转染48 h后,细胞迁移和侵袭能力下降(t=3.47,P<0.05;t=5.72,P<0.01)。细胞凋亡指数(AI)明显增加(t=6.37,P<0.05)。Survivin mRNA和蛋白表达明显下调(t=3.50,P<0.05;t=7.81,P<0.01),Smac mRNA和蛋白表达上调(t=-2.80,P<0.05;t=11.39,P<0.01),VEGF mRNA和蛋白表达明显下调(t=2.54,P<0.05;t=33.84,P<0.01)。结论:ASODN可诱导卵巢癌细胞凋亡,降低卵巢癌细胞侵袭和迁移能力。这些作用是通过下调survivin、VEGF基因,上调Smac基因表达来共同完成的,他们之间的相互作用可能是卵巢癌发生、发展和转移的重要分子机制,survivin可能在其中起关键性作用。针对survivin的基因治疗将成为卵巢癌治疗的重要手段,有重要的临床价值。

Effects of antisense oligonucleotides targeting survivin on apoptosis,invasion and migration of ovarian cancer cells

Objective:To study the effects of antisense oligonucleotides(ASODN)targeting survivin on apoptosis,invasion,and migration of ovarian cancer cell line SKOV3 so as to explore the molecular mechanisms and clinical value of ASODN.Methods:Survivin ASODN was transfected into SKOV3 cells mediated by LipofectamineTM2000.The rate of transfection was detected by flow cytometry and confocal microscopy(SLM510).The invasion and migration abilities were detected by Transwell migration assay.Apoptosis index(AI) and the changes of survivin,Smac,and vascular endothelial growth factor(VEGF) were detected by flow cytometry.The mRNA levels of survivin,Smac,and VEGF were determined by RT-PCR.Results: ASODN was transfected into SKOV3 cells by LipofectamineTM2000 and the rate of transfection was above 95%.The invasion and migration abilities were significantly reduced at 48 h after transfection with 600 nmol/L ASODN(t=3.47,P<0.05;t=5.72,P<0.01).AI was significantly increased(t=6.37,P<0.05).The mRNA and protein levels of survivin and VEGF were significantly down-regulated(t=3.50,P<0.05;t=7.81,P<0.01;t=2.54,P<0.05;t=33.84,P<0.01).But the mRNA and protein levels of Smac were up-regulated(t=-2.81,P<0.05;t=11.39,P<0.01).Conclusion:Survivin ASODN inducs apoptosis and reducs the invasion and migration abilities of ovarian cancer cell line SKOV3 by down-regulating survivin and VEGF expression and upregulating Smac expression.Their interaction may be have an important molecular mechanisms underlying the oncogenesis,metastasis,and development of ovarian cancer.Survivin plays the crucial role during the process.The survivin-targeted gene therapy would be an important approach for the treatment of ovarian cancer and have significant clinical value.