泛素结合酶E2 A在顺铂诱导的HeLa细胞应激反应中的作用

doi:10.3969/j.issn.1004-616x.2018.06.009

癌变·畸变·突变 ›› 2018, Vol. 30 ›› Issue (6): 463-467.doi: 10.3969/j.issn.1004-616x.2018.06.009

泛素结合酶E2 A在顺铂诱导的HeLa细胞应激反应中的作用

王玉强¹, 李红娟¹, 蒋天靓¹, 曹亦菲¹, 杨军^1,2

1. 杭州师范大学医学院, 浙江杭州 310016;
2. 浙江大学附属第一医院传染病诊治国家重点实验室, 浙江杭州 310003

收稿日期:2018-04-13 修回日期:2018-10-29 出版日期:2018-11-30 发布日期:2018-11-30
通讯作者: 杨军,E-mail:gastate@zju.edu.cn E-mail:gastate@zju.edu.cn
作者简介:王玉强,E-mail:wangyq0719@qq.com。
基金资助:
国家自然科学基金（81373036）；浙江省自然科学基金（Z19H260002）；浙江省卫生厅基础研究项目（2014KYB184）

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

WANG Yuqiang¹, LI Hongjuan¹, JIANG Tianliang¹, CAO Yifei¹, YANG Jun^1,2

1. Hangzhou Normal University School of Medicine, Hangzhou 310016;
2. National Key Laboratory for the Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University, Hangzhou 310003, Zhejiang, China

Received:2018-04-13 Revised:2018-10-29 Online:2018-11-30 Published:2018-11-30

摘要/Abstract

摘要： 目的：探讨泛素结合酶E2 A（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UBE2A mRNA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UBE2A mRNA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。

关键词: UBE2A, siRNA, 顺铂, 应激反应

Abstract: OBJECTIVE: To investigate the role of ubiquitin-conjugating enzyme E2 A (UBE2A) in cisplatin-induced 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 cisplatin-induced stress response, via inhibition of UBE2A expression and increased sensitivity to the cytotoxic effect of cisplatin.

Key words: UBE2A, siRNA, cisplatin, stress response

中图分类号:

R992

王玉强, 李红娟, 蒋天靓, 曹亦菲, 杨军. 泛素结合酶E2 A在顺铂诱导的HeLa细胞应激反应中的作用[J]. 癌变·畸变·突变, 2018, 30(6): 463-467.

WANG Yuqiang, LI Hongjuan, JIANG Tianliang, CAO Yifei, YANG Jun. Involvement of UBE2A in cisplatin-induced stress response in HeLa cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(6): 463-467.

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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

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