论文部分内容阅读
目的:利用siRNA技术降低细胞分化抑制因子(Id1)在卵巢癌细胞SKOV3、HO-8910中蛋白水平的表达,并探讨其对化疗耐药性的影响。方法:利用Id1-siRNA干扰Id1 RNA水平的表达,采用蛋白印迹实验检测卵巢癌细胞SKOV3、HO-8910中Id1蛋白水平的表达;转染siRNA后,实验细胞的培养液中加入顺铂,部分复孔在加入顺铂的同时各自加入细胞化学通路抑制剂BHA、Sp600125、Necrostain、z-VAD-fmk,培养48小时后分别利用MTT、LDH检测细胞的生存率、死亡率。结果:①卵巢癌细胞株SKOV3、HO-8910中均有Id1蛋白水平的表达。②Id1-siRNA组中Id1蛋白水平的表达比空白对照组、阴性对照组中的表达明显降低。③Id1-siRNA组细胞死亡率明显低于空白对照组、阴性对照组。④z-VAD-fmk组细胞死亡率明显低于顺铂组,其余各组无明显变化。结论:Id1-siRNA干扰Id1 RNA水平的表达,降低Id1蛋白水平的表达,通过调节下游的凋亡通路,能有效降低卵巢癌细胞株对顺铂的耐药性。
OBJECTIVE: To use siRNA technology to reduce the expression of Id1 in ovarian cancer cells SKOV3 and HO-8910, and to explore its effect on chemoresistance. Methods: Id1-siRNA was used to interfere the expression of Id1 RNA. Western blotting was used to detect the expression of Id1 protein in ovarian cancer cells SKOV3 and HO-8910. After transfected with siRNA, cisplatin was added to the culture medium of the experimental cells, Wells were added cisplatin each while adding cytosolic inhibitors BHA, Sp600125, Necrostain, z-VAD-fmk, after 48 hours of culture, MTT, LDH were used to detect cell viability and mortality. Results: ① The expression of Id1 protein in ovarian cancer cell lines SKOV3 and HO-8910. Id1 protein expression in Id1-siRNA group was significantly lower than that in blank control group and negative control group. ③Id1-siRNA group cell death rate was significantly lower than the blank control group, negative control group. ④z-VAD-fmk group cell death rate was significantly lower than the cisplatin group, the rest of the group no significant change. Conclusion: Id1-siRNA interfered the expression of Id1 RNA and decreased the expression of Id1 protein. By regulating the downstream apoptotic pathway, Id1-siRNA can effectively reduce the drug resistance of ovarian cancer cell lines to cisplatin.