泛素结合酶E2 A在顺铂诱导的HeLa细胞应激反应中的作用

Involvement of UBE2A in cisplatin-induced stress response in HeLa cells

目的:探讨泛素结合酶E2A(UBE2A)基因在顺铂(cisplatin)诱导的HeLa细胞应激反应中的作用。方法:RT-PCR检测顺铂作用后UBE2AmRNA在HeLa细胞中的表达水平;利用针对UBE2A的特异性siRNA转染细胞后,采用qPCR检测UBE2AmRNA在HeLa细胞中的表达情况,并应用CCK-8法检测细胞在顺铂作用下的存活率。结果:UBE2A基因在HeLa细胞中表达两种转录本:U-1和U-2,U-2相比U-1缺少了外显子4的部分序列,并且顺铂作用后二者表达水平均升高。针对UBE2A的特异性siRNA可分别降低其对应转录本的表达水平,表达量分别为对照组的27.43%、15.85%。UBE2A表达受到抑制的HeLa细胞在顺铂作用后细胞存活率降低,相比对照组分别下降了14.46%和19.20%,差异均有统计学意义(P均<0.05)。结论:UBE2A可能参与了顺铂诱导的细胞应激反应,抑制其表达可以明显提高HeLa细胞对顺铂细胞毒性的敏感性。

OBJECTIVE:To investigate the role of ubiquitin-conjugating enzyme E2 A(UBE2A)in cisplatininduced stress response in HeLa cells.METHODS:HeLa cells were transiently transfected with UBE2A-specific siRNA,qPCR was then used to measure the expression level of UBE2A mRNA,and cell viability was measured using the CCK-8 assay after cisplatin treatment.RESULTS:Two different transcripts for UBE2A were identified in HeLa cells:U-1 and U-2,in which U-2 lacks a partial sequence of exon 4 compared to U-1,and both were up-regulated by cisplatin treatment.Expression of the two transcripts could be inhibited by siRNA,with an inhibition ratio of 27.43%for U-1 and 15.85%for U-2 compared to the control group,respectively.The inhibition of their expression in HeLa cells resulted in decreased cell viability after cisplatin treatment,with a 14.46%decrease for U-1 and 19.20%for U-2 compared with the cisplatin treatment group,respectively.CONCLUSION:UBE2A might be involved in cisplatininduced stress response,via inhibition of UBE2A expression and increased sensitivity to the cytotoxic effect of cisplatin.