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Hsa-miR-132 affects proliferation and invasion of esophageal carcinoma cells by targeting FOXA1 gene
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Abstract:
Objective:To investigate the effect of hsamiR132 (miR132) on proliferation and invasion of human esophageal carcinoma cells, and further explore its possible mechanism. Methods: pCDNA3.1(+)miR132 plasmid was constructed, and KYSE150 cells stablely overexpressing miR132 were selected through G418 screening. Soft agar colony formation test and Transwell assay were performed to analyze the proliferation and invasion of KYSE150 cells. Expression of miR132 and forkhead box protens A1 (FOXA1) in KYSE150 cells were examined by realtime PCR and Western blotting. The regulatory effect of miR132 overexpression on FOXA1 was further studied by reporter gene assay and Western blotting. Results: pCDNA3.1(+)miR132 plasmid was successfully constructed, and KYSE150 cells stablely overexpressing miR132 was established. In parental KYSE150 cells, miR132 expression was low ,while FOXA1 expression was high. Compared to mock transfected KYSE150 cells and untransfected KYSE150 cells, pCDNA3.1(+)miR132 transfection did not affect the proliferation of KYSE150 cells (13.9±0.33 vs 15.4±0.11, 17.1±0.20, P>0.05), however tumor size reduced and tumor appeared smoother after pCDNA3.1(+)miR132 transfection. Furthermore, overexpression of miR132 significantly suppressed the invasion of KYSE150 cells（55±1.6 vs 129.0±3.1, 124.0±2.8, P＜001), and miR132 decreased the expression of endogenous FOXA1 by targeting FOXA1 3'UTR. Conclusion: Esophageal carcinoma KYSE150 cells express low level of miR132, and overexpression of miR132 can negatively regulate the expression of FOXA1 and suppressed the invasion of KYSE150 cells.
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Project Supported:
Project supported by the Key Project of Health Bureau of Hebei Province（No. 20110068）
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