外科理论与实践2017，Vol.22Issue(1)：49-56,8.DOI:10.16139/j.1007-9610.2017.01.011
microRNA-130b促进三阴性乳腺癌细胞及其作用机制
microRNA-130b promotes triple-negative breast cancer cell and mechanism
金薇 1罗祎 1刘溦薇 1陈国庆 1周鸣1
作者信息
- 1. 上海交通大学医学院附属同仁医院普外科，上海 200336
- 折叠
摘要
Abstract
Objective To explore the effect of microRNA-130b (miR-130b) expression on proliferation, invasion and migration of triple-negative breast cancer (TNBC) cell and the mechanism. Methods TNBC cell lines MDA-MB-231 were transiently transfected with miR-130b inhibitor, mimic or the negative control (NC) by LipofectamineTM 2000. Successful transfection was confirmed by quantitative real time polymerase chain reaction (qRT-PCR). The function of MDA-MB-231 cells was investigated by the assays of MTT, colony formation, scratch and transwell test after transfection with miR-130b inhibitor mimic and NC. The bioinformatics software was used to look for the potential target gene of miR-130b. Dual-lu-ciferase reporter gene assay was used to verify whether miR-130b was bound to 3′untranslated regions (3′UTR) of potential target gene and the effect was further verified by qRT-PCR and Western blot analysis. Results MDA-MB-231 cells were transfected with miR-130b inhibitor, mimic and NC. The expression of miR-130b increased significantly in mimic group and decreased in inhibitor group when compared with those in NC group with statistical difference significantly (P<0.01). The promotion of cell proliferation (P<0.05), colony formation (P<0.05), invasion (P<0.01) and migration (scratch assay P<0.05, transwell test P<0.05) was promoted in mimic group, whereas down-regulated expression of miR-130b suppressed the ability of proliferation (P<0.05), colony formation (P<0.05), migration (scratch assay P<0.05, transwell test P<0.01) and invasion (P<0.01) when compared with the control cells. According to prediction, CYLD is a potential target of miR-130b based on the prediction with bioinformatics software. Dual-luciferase reporter gene assay revealed that luciferase activity was down-regulated 57.21% in mimic group of transfection with miR-130b mimic +psciCHECK2-CYLD 3′UTR (wild) compared with that in negative group of transfection with miR-NC+psci CHECK2-CYLD 3′UTR (wild) (P<0.001). The ex-pression of CYLD mRNA did not change significantly after the transfection of MDA-MB-231 cells with miR-130b both mimic and inhibitor when compared with that in NC group (P>0.05). However, the expression of CYLD protein levels in-creased in miR-130b inhibitor group and decreased in miR-130b mimic group compared with that in NC group (P<0.001). Conclusions miR-130b could promote proliferation, migration and invasion of TNBC cell lines MDA-MB-231. The effect of miR-130b on TNBC would be achieved partly at least through inhibiting the expression of CYLD.关键词
miR-130b/CYLD/三阴性乳腺癌/增殖/侵袭Key words
miR-130b/CYLD/Triple-negative breast cancer/Proliferation/Invasion分类
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金薇,罗祎,刘溦薇,陈国庆,周鸣..microRNA-130b促进三阴性乳腺癌细胞及其作用机制[J].外科理论与实践,2017,22(1):49-56,8.
