反义硫代寡核苷酸对786-O细胞survivin基因的表达及生长抑制作用

目的研究survivin反义寡核苷酸对肾透明细胞癌786O细胞表达survivin蛋白、细胞凋亡、增殖的影响。方法设计并合成特异性靶向survivin的反义寡核苷酸(ASODN)。肾透明细胞癌细胞株786O分为6组:空白对照组、脂质体转染对照组、正义链转染对照组、200、400和600nmol/LASODN转染组。作用24h后收获各组细胞。倒置显微镜及透射电子显微镜观察细胞形态变化,免疫组化法检测各组细胞survivin表达情况和流式细胞术检测各组细胞周期变化和凋亡指数,MTT法检测survivin反义寡核苷酸对各组细胞的生长抑制率。结果电镜下可以见到典型凋亡样改变,而各对照组细胞生长良好;各ASODN转染组细胞survivin表达有不同程度减弱;各ASODN转染组细胞凋亡指数明显高于各对照组(P<0.05),以600nmol/LASODN转染组最为明显(P<0.05),而各对照组间差异无显著性(P>0.05)。结论不同浓度survivinASODN转染后能下调survivin蛋白表达,诱导肾透明细胞癌786O细胞凋亡,抑制786O细胞增殖。

Antisense oligonucleotide targeting survivin inducing apoptosis of renal cancer cell line 786-O in vitro

Objective To investigate the effect of antisense oligonucleotide (ASODN) targeting survivin on the apoptosis and proliferation of renal cancer cell line 786-O. Methods ASODN targeting survivin was designed and constructed.Cultured cells were divided into 6 groups:control group,liposome group, sense oligonucleotide (SODN) group,200 nmol/L ASODN group,400 nmol/L ASODN group and 600 nmol/L ASODN group.Cells were transfected for 24 h and then harvested to carry on the next tests.Morphological changes of cells were observed by transmission electron microscope. Survivin protein expression was detected by immunohistochemical method and imaging analysis. Apoptosis index (AI) and cell cycle were examined by flow cytometry. Inhibition ratio (IR) was determined by the colorimetri MTT cell viability and proliferation assay. Results Abnormal morphological changes of cells were observed in ASODN transfected groups.Expression of survivin in ASODN groups was significantly decreased compared with control group,liposome group and SODN group.The AI of ASODN groups was significantly higher than that of other groups.The IR of ASODN groups was significantly higher than that of other groups. Conclusion Expression of survivin may decrease 786-O cells after ASODN transfection.ASODN targeting survivin can induce 786-O cell apoptosis and inhibit 786-O cell proliferation.