miR-433-3p Regulates the Biological Behavior of Lung Cancer Cells by Down-Regulating GALNT1
LU Songmei, LEI Shuangyi*, LI Dairong, WANG Yiming, HUANG Lumi, LIANG Guanzhong, YANG Dan
To explore the effects of miR-433-3p (miRNA-433-3p) on proliferation, migration, invasion and apoptosis of lung cancer cells and its molecular mechanism, human normal lung epithelial cells BEAS-2B and lung cancer cell lines H1299, Calu-3 and Calu-6 were cultured in vitro. The RT-qPCR (quantitative reverse transcription) was used to detect the expression of miR-433-3p and GALNT1 (N-acetylgalactosamine transferase 1) mRNA in the above cells; Western blot was used to detect the GALNT1 protein in the above cells. The lung cancer cell which had relatively high expression of miR-433-3p and/or GALNT1 mRNA and protein were selected into the study. The selected cells were divided into the miR-NC group (blank transfection control group), miR-433- 3p group (miR-433-3p overexpression group), si-NC group (blank silent control), si-GALNT1 group (GALNT1 silence group), miR-433-3p+pcDNA group (miR-433-3p and pcDNA co-transfection group), and miR-433- 3p+pcDNA-GALNT1 group (miR-433-3p and pcDNA, pcDNA-GALNT1 co-transfection group). The MTT, Transwell, flow cytometry and Western blot assay were used to detect the cell proliferation, migration, invasion, apoptosis and related protein expression. Dual luciferase reporter assay was used to detect the targeting relationship between miR-433-3p and GALNT1. In this study, it was found that compared with BEAS-2B, the expression level of miR-433-3p in lung cancer cells was significantly lower(P<0.05), and the expression of GALNT1 was significantly increased (P<0.05). Overexpression of miR-433-3p or interference with GALNT1 significantly reduced the process of lung cancer H1299 cells. The expression levels of CyclinD1, MMP-2, MMP-9 and Bcl-2 were significantly reduced (P<0.05), and the expression levels of p21 and Bax were significantly increased (P<0.05). miR-433-3p targeted the expression of GALNT1 (P<0.05). The up-regulation of GALNT1 reversed the effects of miR-433-3p overexpression on the proliferation migration, invasion and apoptosis of lung cancer H1299 cells (P<0.05). In this study, it concluds that miR-433-3p regulates proliferation, migration, invasion and apoptosis of lung cancer H1299 cells by down-regulating the expression of GALNT1.
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