miR-200c对三阴性乳腺癌上皮间质转化的抑制作用及其机制

doi:10.13481/j.1671-587x.20180120

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (01): 106-110.doi: 10.13481/j.1671-587x.20180120

miR-200c对三阴性乳腺癌上皮间质转化的抑制作用及其机制

马啸天¹, 岳小欣², 荣守华¹, 张玉超¹, 田远¹, 石瑛¹, 李君芳³, 贾莉婷¹

1. 郑州大学第三附属医院检验科, 河南郑州 450052;
2. 河南省高等医学专科学校药学系, 河南郑州 451191;
3. 河南省漯河市中心医院, 河南漯河 462000

收稿日期:2017-08-14 出版日期:2018-01-28 发布日期:2018-01-24
通讯作者: 贾莉婷,教授,硕士研究生导师(Tel:0371-66903509,E-mail:jialt@163.com) E-mail:jialt@163.com
作者简介:马啸天(1992-),男,河南省郑州市人,在读医学硕士,主要从事临床免疫学方面的研究。
基金资助:
河南省科技厅医学科技攻关计划重点项目资助课题（201402021）

Inhibitory effect of miR-200c on epithelial-mesenchymal transiton in triple negative breast cancer and its mechanism

MA Xiaotian¹, YUE Xiaoxin², RONG Shouhua¹, ZHANG Yuchao¹, TIAN Yuan¹, SHI Ying¹, LI Junfang³, JIA Liting¹

1. Department of Clinical Laboratory, Third Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China;
2. Department of Pharmacy, Henan Medical College, Zhengzhou 451191, China;
3. Luohe Central Hospital, Henan Province, Luohe 462000, China

Received:2017-08-14 Online:2018-01-28 Published:2018-01-24

摘要/Abstract

摘要： 目的：研究miR-200c对乳腺癌MDA-MB-231细胞和BT-549细胞迁移能力及增殖能力的影响，阐明miR-200c抑制三阴性乳腺癌上皮间质转化（EMT）的相关机制。方法：选取三阴性乳腺癌细胞系MDA-MB-231和BT-549，对其进行瞬时转染，分为空白对照组、阴性对照组（转染negative control/Lipo2000）、试剂对照组（转染Lipo2000）和实验组（转染miR-200c mimic/Lipo2000）；利用RT-PCR和Western blotting法分别检测vimentin及β-catenin mRNA和蛋白表达水平；采用CCK8实验和划痕实验分别检测细胞增殖能力和迁移能力。结果：RT-PCR法和Westernblotting法检测，实验组细胞中vimentin和β-catenin mRNA和蛋白表达水平降低，与空白对照组、阴性对照组和试剂对照组比较差异均有统计学意义（P<0.05）。CCK8法检测，实验组细胞增殖低于阴性对照组和试剂对照组（P<0.05）。划痕实验，实验组细胞划痕宽度恢复率低于阴性对照组和试剂对照组（P<0.05）。结论：miR-200c通过调节MDA-MB-231和BT-549中β-catenin、vimentin的mRNA和蛋白表达量、降低细胞的迁移和增殖能力，进而抑制三阴性乳腺癌EMT。

关键词: β-链接蛋白, 三阴性乳腺癌, 乳腺肿瘤, 波形蛋白, 上皮间质转化, miR-200c

Abstract: Objective: To study the effect of miR-200c on the migration and proliferation abilities of breast cancer MDA-MB-231 and BT-549 cells,and to clarify the mechanism of miR-200c in inhibiting the epithelial-mesenchymal transiton (EMT) of triple negative breast cancer. Methods: The human triple negative breast cancer cell lines (MDA-MB-231 and BT-549) were chosen in this study.The cells were transiently transfected with miR-200c mimics and Lipo2000(experimental group),miR-200c negative control and Lipo2000(negative control group),and Lipo2000 alone (reagent control group); at the same time, blank control group was set up.The expression levels of vimentin and β-catenin mRNA and protein were detected by RT-PCR and Western blotting method.The proliferation rates and migration abilities of MDA-MB-231 cells and BT-549 cells after transfection of miR-200c were analyzed by CCK8 assay and wound healing assay. Results: The RT-PCR and Western blotting showed that the expression levels of vimentin and β-catenin mRNA and proteins in experimental group were decreased,and the differences were statistically significant compared with blank control group, negative control group and reagent control group(P<0.05).The CCK8 results showed that the proliferation rates of the cells in experimental group were lower than those in negative control group and reagent control group(P<0.05).The wound healing assay results showed that the recovery rate of scratch width in experimental group was lower than those in negative control group and reagent control group (P<0.05). Conclusion: miR-200c might inhibit EMT in triple negative breast cancer by regulating the expressions of β-catenin and vimentin mRNA and proteins in MDA-MB-231 and BT-549 cells and decreasing the abilities of migration and proliferation of triple negative breast cancer cells.

Key words: β-catenin, epithelial-mesenchymal transiton, triple negative breast cancer, vimentin, breast neoplasms, miR-200c

中图分类号:

R737.9

马啸天, 岳小欣, 荣守华, 张玉超, 田远, 石瑛, 李君芳, 贾莉婷. miR-200c对三阴性乳腺癌上皮间质转化的抑制作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(01): 106-110.

MA Xiaotian, YUE Xiaoxin, RONG Shouhua, ZHANG Yuchao, TIAN Yuan, SHI Ying, LI Junfang, JIA Liting. Inhibitory effect of miR-200c on epithelial-mesenchymal transiton in triple negative breast cancer and its mechanism[J]. Journal of Jilin University Medicine Edition, 2018, 44(01): 106-110.

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miR-200c对三阴性乳腺癌上皮间质转化的抑制作用及其机制

Inhibitory effect of miR-200c on epithelial-mesenchymal transiton in triple negative breast cancer and its mechanism

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