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目的研究探讨沉默聚腺苷酸二磷酸核糖聚合酶(PARP-1)后,卵巢癌上皮细胞对顺铂药物的敏感性及相关生物学行为的变化。方法以人卵巢癌细胞株A2780为研究对象,采用小RNA干扰技术,沉默PARP-1的表达后,采用Western blot实验、RT-PCR实验、MTT实验等观察PARP-1蛋白、mRNA的表达及细胞存活率的变化趋势,采用方差分析进行比较。结果 PARP1小RNA干扰序列(PARP1-siRNA)转染组PARP-1 mRNA表达量明显低于A2780组和阴性对照组接种转染组(NC-siRNA),3组PARP-1 mRNA表达量分别为(45.22±3.99)%、(24.13±5.24)%、(46.28±6.19)%(P<0.05);PARP1-siRNA转染组PARP-1蛋白表达IOD值明显低于A2780组和NC-siRNA转染组(P<0.05);与0μmol/L相比,5μmol/L和10μmol/L组的PARP-1蛋白表达IOD值明显降低(P<0.05);随着顺铂浓度的增高,PARP1-siRNA转染组的细胞存活率下降的速度最快。结论 PARP1-siRNA转染明显下调人卵巢癌细胞株A2780的PARP-1表达后,该细胞对顺铂药物的敏感性增强,细胞的存活率明显下降。
Objective To investigate the changes of sensitivity and related biological behavior of ovarian cancer epithelial cells to cisplatin after silencing poly (ADP-ribose) polymerase (PARP-1). Methods Human ovarian cancer cell line A2780 was used as a research object. The expression of PARP-1 was silenced by small RNA interference (RNAi) technique. Western blot, RT-PCR and MTT were used to observe the expression of PARP-1 protein and mRNA Survival rate trends, using analysis of variance to compare. Results The expression of PARP-1 mRNA in PARP1-siRNA transfected group was significantly lower than that in A2780 group and negative control group (NC-siRNA). The expressions of PARP-1 mRNA in the three groups were ( 45.22 ± 3.99)%, (24.13 ± 5.24)% and (46.28 ± 6.19)% respectively (P <0.05). The PARP1 protein expression in PARP1-siRNA transfected group was significantly lower than that in A2780 group and NC-siRNA transfected group (P <0.05). Compared with 0μmol / L, PARP-1 protein expression in 5μmol / L and 10μmol / L groups was significantly decreased (P <0.05); PARP1-siRNA was transfected with cisplatin Group cell survival rate dropped the fastest. CONCLUSIONS: PARP1-siRNA transfection significantly down-regulates the expression of PARP-1 in human ovarian cancer cell line A2780, and its sensitivity to cisplatin is enhanced. The cell survival rate is significantly decreased.