Expression of microRNA-380-5p in cervical cancer tissues and cell lines and its inhibition on proliferation and migration of C33Acells by down-regulating RHOA
10.3872/j.issn.1007-385x.2019.01.014
- VernacularTitle:微小RNA-380-5p在宫颈癌组织及细胞中的表达及其通过下调RHOA抑制C33A 细胞的增殖和迁移
- Author:
HAN Weizhen
1
;
JIANG Kun
1
;
YANG Changqun
1
;
YAN Lin
1
;
TAN Zhaoping
1
;
XIONG Guoping
1
Author Information
1. (Department of Obstetrics and Gynecology, the Central Hospital of Wuhan Affiliated to Tongji Medical College, Huazhong University of Science and Technology
- Publication Type:Journal Article
- Keywords:
microRNA-380-5p;
cervical cancer;
RHOA;
proliferation;
migration
- From:
Chinese Journal of Cancer Biotherapy
2019;26(1):85-89
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the expression of microRNA-380-5p (miR-380-5p) in cervical cancer tissues and cell lines, and to explore the mechanism of miR-380-5p inhibiting the proliferation and migration of cervical cancer cells. Methods: 16 pairs of cervical cancerous tissues and corresponding para-cancerous tissues were collected from the Department of Obstetrics and Gynecology, the Affiliated Wuhan Central Hospital of Tongji Medical College from December 2016 to July 2017; in addition, cervical cancer cell lines (HCC94, C33A, Hela, SiHa) and human cervical epithelial immortalized H8 cells were also collected for this study. The expression of miR-380-5p in above mentioned tissues and cell lines was detected by Real-time quantitative polymerase chain reaction (qPCR). miR380-5p mimic (experimental group) and miR-NC (negative control group) were transiently transfected into C33A cells by lipofection, and qPCR was used to detect the expression of miR-380-5p in the transfected cells. Cell proliferation and migration were evaluated by cell counting kit (CCK-8) and Transwell assay. Bioinformatics software TargetScan predicted the downstream genes of miR-380-5p, and dual luciferase reporter assay was used to verify the binding of miR-380-5p to the downstream gene RHOA (Ras homolog gene family member A). qPCR and Western blotting were used to detect the expression of miR-380-5p downstream gene-RHOA. Results: The expression level of miR-380-5p in cervical cancer tissues and cell lines was significantly lower than that in para-cancerous tissues and normal cervical epithelial H8 cells (P<0.01); and the expression in C33A cells was the lowest (P<0.01). Compared with the negative control group, the miR-380-5p mimic transfection singnificantly inhibited the proliferation (P<0.05) and migration ability of C33A cells (P<0.01), and down-regulated protein expressions of RHOA, ROCK1, ROCK2, CDK2 and N-cadherin (all P<0.01). Bioinformatics software predicted that RHOA may be a downstream gene of miR-380-5p, and dual luciferase reporter assay proved the specific binding of miR-380-5p to the 3'UTR of RHOA (P<0.01). miR-380-5p could significantly down-regulate RHOA gene expression (P< 0.01). Conclusion: miR-380-5p is low-expressed in cervical cancer cell lines. Over-expression of miR-380-5p may inhibit the proliferation and migration of cervical cancer C33Acells by down-regulating the expression of RHOAgene and its downstream proteins.
- Full text:20190114.pdf
