论文部分内容阅读
目的:运用RNA干扰技术双向沉默卵巢癌耐顺铂细胞株A2780-cp的Survivin和XIAP基因后,观察细胞的凋亡及对顺铂敏感性的影响。方法:通过脂质体介导将化学合成的Survivin-siRNA和XIAP-siRNA,共同或单独转染到卵巢癌耐药细胞株A2780-cp中,未转染组作为阴性对照;流式细胞术(FCM)检测转染细胞凋亡率,蛋白质印迹法检测Survivin和XIAP基因的蛋白表达,四甲基偶氮唑盐(MTT)法检测转染后细胞对顺铂的敏感性及其增殖情况。结果:转染48 h后,单独和共同转染组与对照组相比可见明显的细胞凋亡形态学变化,共转染组凋亡率为26.98%,高于单转Survivin-siRNA组(12.82%)和单转XIAP-siRNA组(11.18%)。蛋白质印迹法结果显示,共转染组Survivin和XIAP的蛋白抑制率分别为71.11%和76.81%,单转染组Survivin和XIAP蛋白抑制率分别为75.00%和82.48%,与其他对照组比较差异有统计学意义,P<0.05;MTT结果显示,转染后细胞对顺铂敏感性增强,共转染组细胞生长活力下降明显,顺铂的IC50值为(0.10±0.01)μg/mL,与单转染组及脂质体转染组比较差异有统计学意义,P<0.05。结论:同时干扰Survivin基因和XIAP基因能更好的抑制卵巢癌耐药细胞株A2780-cp的生长,降低其耐药性,促进其凋亡。
OBJECTIVE: To investigate the effects of RNA interference on the apoptosis and the sensitivity of cisplatin to cisplatin-resistant ovarian cancer A2780-cp cell line A2780-cp. Methods: Chemically synthesized Survivin-siRNA and XIAP-siRNA were transfected into A2780-cp ovarian cancer cell line by either lipofectamine or non-transfected cells. Flow cytometry FCM was used to detect the apoptosis rate of transfected cells. Western blotting was used to detect the protein expression of Survivin and XIAP. MTT was used to detect the sensitivity and proliferation of transfected cells to cisplatin. Results: After 48 h of transfection, the apoptotic morphological changes were obvious compared with the control group. The apoptosis rate of transfected group was 26.98%, higher than that of Survivin-siRNA group (12.82 %) And single XIAP-siRNA group (11.18%). The results of Western blotting showed that the inhibitory rates of Survivin and XIAP were 71.11% and 76.81%, respectively. The inhibitory rates of Survivin and XIAP in single transfection group were 75.00% and 82.48%, respectively. Compared with other control groups, there were (P <0.05). The results of MTT showed that the transfected cells had a higher sensitivity to cisplatin and the cell viability decreased significantly in the co-transfected cells. The IC50 of cisplatin was (0.10 ± 0.01) μg / mL, There was significant difference between transfection group and lipofection group (P <0.05). Conclusion: Simultaneous interference of Survivin gene and XIAP gene can better inhibit the growth of drug-resistant A2780-cp ovarian cancer cell line, reduce its drug resistance and promote apoptosis.