三阴性乳腺癌MDA-MB-231细胞低氧模型的建立及作用研究

doi:10.3969/j.issn.1004-616x.2016.06.002

癌变·畸变·突变 ›› 2016, Vol. 28 ›› Issue (6): 420-427.doi: 10.3969/j.issn.1004-616x.2016.06.002

三阴性乳腺癌MDA-MB-231细胞低氧模型的建立及作用研究

刘丽桃^1,2, 刘文兰², 万丽丽², 朱婷^1,2, 王国民³, 阮海英², 谢妮²

1. 广州医科大学研究生院, 广东广州 511436;
2. 深圳市第二人民医院, 广东深圳 518037;
3. 深圳肖传国医院, 广东深圳 518129

收稿日期:2016-04-13 修回日期:2016-08-16 出版日期:2016-11-30 发布日期:2016-11-30
通讯作者: 谢妮,E-mail:kejiaoke100@163.com E-mail:kejiaoke100@163.com
作者简介:刘丽桃,E-mail:liulitao1985@yeah.net。
基金资助:
广东省科技计划项目（2013B021800097；2014A020212038）；深圳市知识创新计划基础研究项目（JCYJ20150330102720122）；广东省自然科学基金（2016A030313029）

Impact of different hypoxia conditions on the MDA-MB-231 triple-negative human breast cancer cells

LIU Litao^1,2, LIU Wenlan², WAN Lili², ZHU Ting^1,2, WANG Guomin³, RUAN Haiying², XIE Ni²

1. Graduate School of Guangzhou Medical University, Guangzhou 511436;
2. Second People's Hospital, Shenzhen 518037;
3. Shenzhen C. G. Xiao Hospital, Shenzhen 518129, Guangdong, China

Received:2016-04-13 Revised:2016-08-16 Online:2016-11-30 Published:2016-11-30

摘要/Abstract

摘要：

目的：研究不同的低氧环境对三阴性乳腺癌细胞株MDA-MB-231增殖和侵袭迁移能力的影响。方法：将MDA-MB-231细胞分为间歇低氧组、持续低氧组和常氧组，采用划痕法和Transwell小室法进行迁移实验；CCK8法检测细胞增殖；荧光定量PCR和Western blot分别检测低氧诱导因子-1α（HIF-1α）和波形蛋白（vimentin）的mRNA和蛋白表达；siRNA干扰间歇低氧组细胞HIF-1α基因表达，在此基础上进一步检测低氧细胞的迁移、增殖及vimentin蛋白改变。结果：间歇低氧促进MDA-MB-231细胞迁移，且其促进作用显著高于持续低氧组（P < 0.05）；间歇低氧和持续低氧均抑制细胞增殖，但持续低氧对细胞增殖抑制的作用在低氧48 h后才较为明显（P < 0.05）；间歇性低氧组细胞HIF-1α和vimentin蛋白表达均显著高于持续低氧细胞。siRNA干扰间歇低氧组细胞HIF-1α表达后，其vimentin蛋白下调，细胞迁移能力下降（P<0.05），但增殖水平未受到明显影响（P > 0.05）。结论：间歇低氧可诱导高侵袭表型的MDA-MB-231细胞株，其侵袭力增强与HIF-1α表达增加进而上调vimentin有关。

关键词: 低氧, HIF-1α, 三阴性乳腺癌, 细胞迁移

Abstract:

OBJECTIVE: To study the influence of hypoxia on metastatic potential of the MDA-MB-231 triplenegative human breast cancer cells. METHODS: MDA-MB-231 cells were divided into three hypoxia groups:intermittent hypoxia (IH),continuous hypoxia (CH) and normoxic (N). Cell migration and invasion ability were analyzed by wound healing and the Boyden chamber assay. Cell proliferation was analyzed by the CCK-8 assay. Hypoxia-inducible factor-1α (HIF-1α) and vimentin expression in response to different hypoxic environments were analyzed by Western blot and real-time PCR assays. HIF-1α expression via siRNA knockout was investigated. RESULTS: IH-treated cells exhibited higher invasiveness than the C H-treated cells (P < 0.05). On the other hand,IH significantly inhibited cell proliferation while CH did not show such effect until 48 h later (P < 0.05). IH induced a greater effect on HIF-1α protein accumulation and vimentin upregulation. Knockdown of HIF-1α by siRNA abolished IH-induced cell migration and vimentin upregulation (P < 0.05). However, knockdown of HIF-1α had no effect on proliferation of MDA-MB-231 cells (P > 0.05). CONCLUSION: IH had a more pronounced effect one nhancing the invasive phenotype of M DA-MB-231 cells than C H,and H IF-1α a ctivationt ogether with increased vimentin upregulation might be responsible for the phenotypic change.

Key words: hypoxia, HIF-1α, triple-negative breast cancers, cell migration

中图分类号:

R73-37

刘丽桃, 刘文兰, 万丽丽, 朱婷, 王国民, 阮海英, 谢妮. 三阴性乳腺癌MDA-MB-231细胞低氧模型的建立及作用研究[J]. 癌变·畸变·突变, 2016, 28(6): 420-427.

LIU Litao, LIU Wenlan, WAN Lili, ZHU Ting, WANG Guomin, RUAN Haiying, XIE Ni. Impact of different hypoxia conditions on the MDA-MB-231 triple-negative human breast cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(6): 420-427.

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三阴性乳腺癌MDA-MB-231细胞低氧模型的建立及作用研究

Impact of different hypoxia conditions on the MDA-MB-231 triple-negative human breast cancer cells

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