上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

抑癌基因VHL在H2O2诱导的肿瘤细胞氧化应激中的作用

李彩霞1，毛玉琴1，高耀辉2，韩三峰2，孟 超3，王立顺1   

  1. 1.上海交通大学 医学院附属瑞金医院分子医学中心， 上海 200025； 2.上海市闵行区中心医院， 上海 201100; 3.上海交通大学 医学院附属仁济医院老年科, 上海 200001
  • 出版日期:2014-10-28 发布日期:2014-10-28
  • 通讯作者: 王立顺, 电子信箱: jywangls@shsmu.edu.cn。
  • 作者简介:李彩霞（1987—）， 女， 住院医师， 硕士; 电子信箱: licaixiaowen@163.com。
  • 基金资助:

    国家自然科学基金（31170783）；上海市自然科学基金（11ZR1421500）

Effects of tumor suppressor gene VHL on H2O2 induced oxidative stress  of tumor cells

LI Cai-xia1, MAO Yu-qin1, GAO Yao-hui2, HAN San-feng2, MENG Chao3, WANG Li-shun1   

  1. 1.Center of Molecular Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; 2.Central Hospital of Shanghai Minhang District, Shanghai 201100, China; 3.Department of Gerontology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200001, China
  • Online:2014-10-28 Published:2014-10-28
  • Supported by:

    National Natural Science Foundation of China，31170783; Natural Science Foundation of Shanghai，11ZR1421500

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摘要：

目的 观察抑癌基因VHL对H2O2诱导的肿瘤细胞氧化应激的影响，并初步探究其分子机制。方法 采用RNAi沉默秀丽线虫vhl-1和肿瘤细胞的同源基因VHL的表达，以不同浓度H2O2处理，经形态学分析秀丽线虫的死亡情况；用AnnexinⅤ和PI标记细胞，经流式细胞仪分析细胞死亡情况；采用Western blotting检测氧化应激的通路蛋白c-Jun氨基末端激酶（JNK）的磷酸化水平，分析VHL影响细胞氧化应激的分子机制。结果 与对照组比较，vhl-1表达沉默的秀丽线虫对H2O2的敏感性显著增加，死亡率显著增高，该效应依赖于H2O2的浓度梯度和处理时间。沉默VHL表达肿瘤细胞的死亡率与野生型肿瘤细胞比较无明显差别，但经250 μmol/L和500 μmol/L的H2O2处理后，沉默VHL表达肿瘤细胞的死亡率显著增高，且伴随JNK磷酸化水平的升高。结论 沉默VHL表达可增加细胞对H2O2诱导的氧化应激的敏感性，且VHL对细胞在H2O2诱导的氧化应激中的保护效应存在种属的保守性。

关键词: 氧化应激, VHL, c-Jun氨基末端激酶

Abstract:

Objective To observe the effects of tumor suppressor gene VHL on the H2O2 induced oxidative stress of tumor cells and to primarily explore the molecular mechanism. Methods The expressions of vhl-1 gene of C.elegans and homologous gene VHL of cancer cells were silenced by RNAi. The death of C.elegans was morphologically analyzed after being treated by H2O2 of different concentrations. Cells were labeled by AnnexinⅤ and PI and the death of cells was analyzed by the flow cytometry. Then the phosphorylation level of pathway protein c-Jun N-terminal kinase (JNK) of oxidative stress was detected by the Western blotting and the molecular mechanism of effects of VHL on the oxidative stress of cells was analyzed. Results Compared to the control group, the sensitivity of C.elegans with silenced VHL gene towards H2O2 increased significantly and the mortality rate also increased significantly. These effects depended on the concentration gradient and treatment time of H2O2. The mortality rate of tumor cells with silenced VHL gene was not significantly different from that of wild type tumor cells. But the mortality rate of tumor cells with silenced VHL gene significantly increased and the phosphorylation level of JNK also increased after being treated by H2O2 of 250 μmol/L and 500 μmol/L. Conclusion Silencing the expression of VHL can increase the sensitivity of cells towards the oxidative stress induced by H2O2. The protective effect of VHL on the H2O2 induced oxidative stress is conserved in species.

Key words: oxidative stress, VHL, c-Jun N-terminal kinase