PINK1在人卵巢癌细胞系SKOV3增殖 及顺铂诱导细胞死亡中的作用

Effect of PINK1 on cell proliferation and cisplatininduced cell deathin human ovarian cancer SKOV3 cells

关键词:  线粒体丝苏氨酸蛋白激酶PINK1  卵巢癌  细胞增殖  顺铂耐药性

Key words:  PINK1  ovarian cancer  proliferation  chemoresistance

摘要:

目的　研究线粒体丝苏氨酸蛋白激酶PINK1（PTEN induced putative kinase 1）在卵巢癌细胞生长增殖中的作用，及其表达在顺铂诱导细胞死亡中的作用。方法　转染并用G418筛选稳定表达pcDNA3.1-PINK1及pcDNA3.1的SKOV3细胞系，细胞设3组：正常培养组、转染空质粒pcDNA3.1组，过表达PINK1组。采用实时定量PCR检测各组细胞PINK1 mRNA的表达，MTT和流式细胞仪检测各组细胞生长增殖及细胞周期分布情况。观察PINK1在顺铂（DDP）诱导卵巢癌细胞死亡中的作用，并检测耐药基因MDR1、MRP1以及ABCG2在各处理组中的表达水平。结果　过表达PINK1的SKOV3细胞系PINK1 mRNA的表达水平显著高于正常培养组及转染空质粒pcDNA3.1组细胞（P<0.05）。过表达PINK1组较其他两组细胞增殖率显著提高，细胞进入S和G₂/M期比例增加（P<0.05）。过表达PINK1显著抑制了10 μmol/L和 20 μmol/L顺铂诱导细胞死亡，并诱导了耐药基因MDR1、MRP1和ABCG2的表达升高。 结论　上调PINK1的表达可促进卵巢癌细胞增殖，降低细胞对顺铂作用的敏感性并诱导耐药基因的表达。

Abstract:

ObjectiveTo observe the effect of mitochondria serine/threonine kinase PINK1 on proliferation of human ovarian cancer SKOV3 cells and its role in cell death during cisplatin exposure.MethodsPINK1 gene stable expression plasmid was transfected to SKOV3 cells, and G418 was used to select successfully transfected cell clones. PINK1,MDR1, MRP1 and ABCG2 expression was detected by real-time qPCR in mRNA level. Cell proliferation ability and cell cycle distributions were analyzed by MTT and flow cytometry.ResultsPINK1 mRNA in PINK1 gene transfected cells was significantly higher than empty plasmid transfected cells and non-transfected cells(P＜0.05).The cell proliferation and the proportion in the S and G₂/M phases were higher in PINK1 gene transfected cells than others(P＜0.05).PINK1 gene transfected cells were more resistant to cisplatin induced cell death, and drug resistantgenes MDR1, MRP1 and ABCG2 expression increased (P＜0.05).ConclusionPINK1 overexpression in SKOV3 cells could decrease cisplatin induced cell death by stimulating drug resistantgenes expression.