整合素连接激酶反义寡核苷酸对卵巢上皮性癌细胞生长的抑制作用
Inhibitory effect of antisense oligonucleotide of integrin-linked kinase on cell proliferation of human epithelial ovarian cancer
目的 探讨整合素连接激酶(ILK)反义寡核苷酸(ASODN)对卵巢上皮性癌(卵巢癌)细胞生长的抑制作用.方法 将ILK-ASODN转染入卵巢癌细胞株H08910细胞,阻断其表达ILK.实验分为6组,分别为空白对照组(A组)、脂质体对照组(B组)、ILK-正义寡核苷酸(SODN)100 nmol/L组(C组)和ILK-ASODN 60 nmol/L组(D组)、ILK-ASODN 80 nmol/L组(E组)、ILK-ASODN 100 nmol/L组(F组).采用RT-PCR技术和蛋白印迹法检测各组H08910细胞ILK mRNA和蛋白表达量的变化;水溶性四氮唑1(WST-1)法及流式细胞术评价H08910细胞转染ILK-ASODN后对该细胞体外增殖的抑制作用及对该细胞周期和凋亡的影响.结果 ILK-ASODN转染后H08910细胞ILK mRNA表达量明显下降,D、E、F 3个实验组分别为0.307±0.011、0.198±0.008、0,与A、B两对照组(分别为0.343±0.006、0.342±0.009)比较,差异均有统计学意义(P<0.05).ILK-ASODN转染后H08910细胞ILK蛋白表达量也明显下降,D、E、F 3个实验组分别为26.3±0.8、20.6±0.4、0;细胞生长受到明显抑制,细胞凋亡率明显增高,3个实验组分别为7.31%、8.84%、11.27%;G0/G1期细胞增多,3个实验组分别为49.25%、56.28%、67.61%.以上指标分别与A、B组比较,差异均有统计学意义(P<0.01).结论 ILK-ASODN转染卵巢癌H08910细胞后可以明显抑制其生长.
更多Objective To explore the inhibitory effect of integrin-linked kinase antisense oligonucleotide(ILK-ASODN)on cell proliferation in human ovarian cancer cell line(HO8910).Methods We transfected ILK-ASODN into HO8910 to block ILK gene expression.measured the expression levels of integirn-linked kinase(ILK)mRNA by RT-PCR and ILK protein by western-blotting;the inhibiting effects of the transfection on HO8910 proliferation,the cell cycles,and cell apoptosis were assessed by water soluble tetrazolium-1(WST-1)and flow cytometry(FCM).Results After transfection of ILK-ASODN,the expression levels of ILK mRNA decreased significantly in groups D,E,F being 0.307±0.011,0.198±0.008,0,respectively,when compared with those of the two control groups of A and B(P<0.05).The expression levels of ILK protein of the groups D,E and F decreased significantly also,being 26.3 ± 0.8,20.6±0.4 and O,respectively.HO8910 cell proliferation was inhibited significantly,and the rates of apoptosis of the groups D,E and F increased significantly,being 7.31%,8.84%and 11.27%respectively.The cell population increased in G0/G1 phase of the groups D,E and F,being 49.25%,56.28%and 67.61%respectively.significantly different in comparison with those of groups of A and B (P<0.01).Conclusions Transfection of ILK-ASODN into human ovarian cancer line inhibited cancer cell proliferation significantly.
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