遗传 ›› 2019, Vol. 41 ›› Issue (4): 318-326.doi: 10.16288/j.yczz.18-278

• 研究报告 • 上一篇    下一篇

CSN4基因干扰对乳腺癌MDA-MB-231细胞增殖和凋亡的影响

余同露1,2,蔡栋梁1,2,朱根凤1,2,叶晓娟1,2,闵太善1,2,陈红岩1,2,卢大儒1,2,陈浩明1,2()   

  1. 1. 复旦大学生命科学学院,上海 200438
    2. 复旦大学现代人类学教育部重点实验室,上海 200438
  • 收稿日期:2018-12-11 修回日期:2019-02-18 出版日期:2019-04-20 发布日期:2019-03-15
  • 通讯作者: 陈浩明 E-mail:hmchen@fudan.edu.cn
  • 作者简介:余同露,硕士研究生;专业方向：生物工程。E-mail: 15210700122@fudan.edu.cn
  • 基金资助:
    国家自然科学基金项目(81071739);国家自然科学基金项目(81272385)

Effects of CSN4 knockdown on proliferation and apoptosis of breast cancer MDA-MB-231 cells

Tonglu YU1,2,Dongliang Cai1,2,Genfeng Zhu1,2,Xiaojuan Ye1,2,Taishan Min1,2,Hongyan Chen1,2,Daru Lu1,2,Haoming Chen1,2()   

  1. 1. School of Life Sciences, Fudan University, Shanghai 200438, China
    2. MOE Key Laboratory of Contemporary Anthropology, Fudan University, Shanghai 200438, China
  • Received:2018-12-11 Revised:2019-02-18 Online:2019-04-20 Published:2019-03-15
  • Contact: Chen Haoming E-mail:hmchen@fudan.edu.cn
  • Supported by:
    the National Natural Science Foundation of China(81071739);the National Natural Science Foundation of China(81272385)

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摘要：

乳腺癌目前是危害女性常见的恶性肿瘤,研究发现COP9复合体中的各个亚基与恶性肿瘤的发生发展密切相关,且CSN4亚基对整个复合体具有很重要的调节作用。为探讨CSN4基因在乳腺癌细胞中的生物学功能及其分子作用机制,本研究首先在乳腺癌细胞MDA-MB-231中,成功建立了慢病毒介导的CSN4基因的干扰表达体系,并通过细胞表型实验、CCK8实验和细胞克隆形成实验证实CSN4基因表达的干扰能显著抑制乳腺癌MDA-MB-231细胞的增殖速率和克隆形成能力。流式细胞检测结果表明,干扰CSN4的表达能显著增加Sub G1期乳腺癌MDA-MB-231细胞比例;凋亡检测结果表明,干扰CSN4的表达能显著增加早期凋亡和晚期凋亡细胞的比例,这证明干扰CSN4的表达能够促进乳腺癌细胞凋亡。最后,通过Western blot实验证实CSN4能调控CDK6和Caspase3蛋白的表达,激活Caspase3切割PARP,揭示CSN4可调控CDK6和Caspase3 的表达从而影响乳腺癌细胞的增殖和凋亡。本研究结果进一步加深了人们对乳腺癌细胞凋亡和细胞生长的分子机制的认识,同时也进一步揭示了CSN4在肿瘤生物学中的作用和机制。

关键词: 乳腺癌, CSN4, 细胞增殖, 细胞凋亡, CDK6, Caspase3

Abstract:

Breast cancer is one of the most common malignant tumors endangering women. It has been found that the subunits of the COP9 complex are closely related to the occurrence and development of malignant tumors, and the CSN4 subunit plays an important role in regulating the whole complex. In the breast cancer cell line MDA-MB-231, we successfully established a lentivirus-mediated CSN4-knockdown cell line. CCK8 cell proliferation assays and colony formation experiments confirmed that CSN4 knockdown significantly decreased the cellular proliferation rate. Cell cycle analysis showed that CSN4 knockdown increased sub-G1 population and induced apoptosis. In addition, Western blotting assays confirmed that CSN4 regulates the expression of CDK6 and Caspase3, suggesting that CSN4 modulates the proliferation and apoptosis of breast cancer cells by regulating the expression of CDK6 and Caspase3 genes and thereby tumorigenesis. This study has deepened our understanding of the molecular mechanism of apoptosis and cell growth in breast cancers, and further revealed the role and mechanism of CSN4 in cancer biology.

Key words: breast cancer, CSN4, cell proliferation, cell apoptosis, CDK6, Caspase3