MicroRNA-133b suppresses cell proliferation and invasion of esophageal squamous cell carcinoma via downregulating TAGLN2 expression
10.3760/cma.j.issn.0253-3766.2019.02.003
- VernacularTitle: miR－133b通过下调转胶蛋白抑制食管鳞癌细胞的增殖和侵袭能力
- Author:
Yu TANG
1
;
Junhao LIU
1
;
Zuxuan SHI
2
;
Zhen LI
2
;
Hongtao LIU
3
;
Ping LU
1
Author Information
1. Department of Endocrinology, Henan Provincial People′s Hospital, Fuwai Central China Cardiovascular Hospital, Zhengzhou 450003, China
2. Department of Medical Oncology, Henan Provincial People′s Hospital, Zhengzhou 450003, China
3. College of Life Sciences, Zhengzhou University, Zhengzhou 450001, China
- Publication Type:Journal Article
- Keywords:
Esophageal neoplasms;
Squamous cell carcinoma;
Proliferation;
Invasion;
Epithelial-mesenchymal transitions;
MiR-133b;
Transgelin-2
- From:
Chinese Journal of Oncology
2019;41(2):91-96
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the expression of microRNA-133b (miR-133b) in esophageal squamous cell carcinoma (ESCC), and explore its effect and the underlying molecular mechanisms on cell proliferation and invasion.
Methods:Real-time quantitative PCR (qPCR) was used to examine miR-133b expression in 63 ESCC tissues and paired adjacent non-cancerous tissues, several ESCC cells (Eca109, EC9706, EC1, TE1, KYSE70) and normal esophageal epithelial cell Het-1A. MiR-133b mimic, inhibitor and negative control (NC) were transfected into TE1 cells. The effect of miR-133b on cell proliferation and invasion were determined by CCK-8 and Transwell assays, respectively. Subsequently, the target gene of miR-133b was predicted by online tools TargetScan and miRDB, which was verified by dual luciferase reporter assays. Finally, Western blot was utilized to detect the effects of miR-133b overexpression on expression of target gene TAGLN2 as well as EMT-related proteins E-cadherin, N-cadherin, Snail, Slug and Vimentin.
Results:Relative levels of miR-133b in ESCC tissues (0.295±0.040) were significantly lower than those in adjacent non-cancerous tissues (1.002±0.011, P<0.001). The expression of miR-133b was tightly associated with clinical staging, lymph node metastasis and prognosis. Moreover, relative levels of miR-133b in ESCC cells Eca109, EC9706, EC1, TE1 and KYSE70 (0.679±0.031, 0.391±0.008, 0.236±0.016, 0.031±0.005 and 0.099±0.020) were evidently lower than that in normal esophageal epithelial cell Het-1A (1.005±0.016, all P<0.001). In TE1 cells, miR-133b mimic significantly increased the level of miR-133b to 6.199±0.627, and suppressed cell proliferation and invasion, whereas miR-133b inhibitor obviously decreased its expression to 0.182±0.023, and promoted cell proliferation and invasion. Most notably, the relative luciferase activities of miR-133b-mimic group (0.320±0.018) in TE1 cells transfected with TAGLN-3′UTR-WT were markedly lower than that in NC group (1.010±0.036, P<0.001), whereas those in TAGLN-3′UTR-MUT transfection cells were 1.019±0.056 and 1.008±0.021, respectively, showing no significantly statistical difference (P>0.05). Furthermore, miR-133b overexpression markedly downregulated TAGLN2, N-cadherin, Snail, Slug and Vimentin levels, and increased E-cadherin expression.
Conclusion:MiR-133b plays an important role in the proliferation and invasion of ESCC cells by regulating TAGLN2 expression, and it may be a potential therapeutic target for ESCC patients.
