mTOR通路激活后上调SNCG表达水平抑制乳腺癌细胞辐射敏感性的研究

doi:10.3969/j.issn.1004-616x.2017.03.009

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (3): 205-210,215.doi: 10.3969/j.issn.1004-616x.2017.03.009

mTOR通路激活后上调SNCG表达水平抑制乳腺癌细胞辐射敏感性的研究

吴丽娜, 高刚, 刘建香

中国疾病预防控制中心辐射防护与核安全医学所辐射防护与核应急中心, 中国疾病预防控制中心重点实验室, 北京 100088

收稿日期:2016-07-12 修回日期:2017-03-11 出版日期:2017-05-31 发布日期:2017-05-31
通讯作者: 刘建香,E-mail:jxliu@nirp.cn E-mail:jxliu@nirp.cn
作者简介:吴丽娜,E-mail:xuanxuan19810123@sohu.com。

Activation of mTOR signaling pathway can reduce radiation sensitivity through up-regulating SNCG expression in breast cancer cells

WU Lina, GAO Gang, LIU Jianxiang

Key Laboratory of Radiological Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, China

Received:2016-07-12 Revised:2017-03-11 Online:2017-05-31 Published:2017-05-31

摘要/Abstract

摘要： 目的：探讨乳腺癌T47D细胞通过激活mTOR通路调控SNCG表达水平，从而抑制乳腺癌细胞辐射敏感性的分子机制。方法：检测不同剂量γ射线照射后的T47D乳腺癌细胞中mTOR蛋白表达水平。在细胞培养液中加入不用浓度磷脂酸（PA，mTOR通路激活剂）进行培养，以常规培养细胞为对照组，采用Western blot法检测对照组和激活剂组细胞SNCG蛋白的表达。对照组和激活组细胞采用4 Gy γ射线照射24 h，检测照射后SNCG mRNA和蛋白的表达情况，并采用平板细胞克隆形成实验检测克隆形成率。同时，将转染SNCG siRNA的乳腺癌T47D细胞株分成激活组和对照组，验证SNCG在乳腺癌细胞辐射敏感性抵抗中的生物学功能。结果：不同剂量射线照射后，mTOR蛋白表达水平显著升高。mTOR激活剂PA处理后的细胞对乳腺癌细胞放射敏感性具有明显的抑制作用，同时Western blot显示γ射线照射处理后的乳腺癌细胞中SNCG蛋白的表达水平异常。Western blot和qPCR方法检测发现，T47D对照组和干扰SNCG基因的T47D细胞实验组中，激活mTOR或γ射线照射均能引起SNCG蛋白和mRNA表达增加。克隆形成实验进一步证明，降低SNCG的表达可显著抑制T47D细胞克隆形成能力。结论：在乳腺癌细胞中，mTOR介导的SNCG表达调控对乳腺癌细胞抗辐射起着重要作用，降低SNCG的表达可提高辐射敏感性，提示在临床治疗中有可能通过使用SNCG抑制剂或mTOR抑制剂提高乳腺癌细胞在放化疗中的敏感性。

关键词: 乳腺癌, mTOR通路, SNCG, 放射治疗

Abstract: OBJECTIVE:To investigate the involvement of mTOR-regulated SNCG expression in the control of radiation sensitivity in T47D breast cancer cells. METHODS:Expression levels of tumor-related genes in T47D cells were investigated after their irradiation with different doses of γ-rays. Western blot analysis was performed to determine the level of SNCG expression with or without the treatments of phosphatidic acid (PA;an mTOR pathway activator),and with 4 Gy γ-rays with or without PA treatment. Furthermore,SNCG expression was inhibited by transfection with SNCG-siRNA,and colony formation assay was performed to document radiation sensitivity. RESULTS:After γ-rays irradiation,there was a remarkable up-regulation of mTOR expression in T47D cells. However,radiation sensitivity was altered in PA-treated cells,suggesting that mTOR pathway was involved in the regulation of radio-sensitivity in these cells. From the qPCR and Western blot analyses,SNCG expression was up-regulated in either γ-rays irradiated or PA-treated T47D cells,indicating that mTOR-mediated the expression of SNCG which reduced the radiation sensitivity in T47D cells. Furthermore,transfection of cells with SNCG-siRNA significantly suppressed colony formation,suggesting that knockdown of SNCG expression can improve radiation sensitivity in breast cancer cells. CONCLUSION:Our study show that mTOR mediated the expression of SNCG and the subsequent radiation sensitivity of breast cancer cells. Our data suggest that down-regulation of SNCG expression may increase the sensitivity of radiotherapy and this may be helpful for the treatment of the patients with breast cancer.

Key words: breast cancer, mTOR pathway, SNCG, irradiation therapy

中图分类号:

R811.5

吴丽娜, 高刚, 刘建香. mTOR通路激活后上调SNCG表达水平抑制乳腺癌细胞辐射敏感性的研究[J]. 癌变·畸变·突变, 2017, 29(3): 205-210,215.

WU Lina, GAO Gang, LIU Jianxiang. Activation of mTOR signaling pathway can reduce radiation sensitivity through up-regulating SNCG expression in breast cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(3): 205-210,215.

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mTOR通路激活后上调SNCG表达水平抑制乳腺癌细胞辐射敏感性的研究

Activation of mTOR signaling pathway can reduce radiation sensitivity through up-regulating SNCG expression in breast cancer cells

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