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- Title
MiR-301b抑制转录因子Klf4 影响肝癌细胞迁移.
- Authors
董笑; 王凡; 刘传; 宋卫峰; 李琦
- Abstract
Objective: To investigate the effect of miR-301b on the migration of hepatocellular carcinoma cells and the underlying molecular mechanisms. Methods: Human liver cancer cell lines including SK-Hep-1, HCC-LM3 and human immortalized liver cell line L02 were cultured in vitro. The expression of miR-301b was detected by RT-PCR. Through bioinformatics software Targetscan and mi-Randa. we predicted target genes of iniR-301b and selected transcription factor K1f4 gene as the downstream target gene of miR-301b. Through the dual luciferase reporter gene experiment and Western Blot experiment, we proved its regulating effect. The role of miR-301b on the migration of hepatocellular carcinoma cells were verified by wound healing assay and transwell migration assay. The effect of miR-301b on the expression of E-cadherin and N-cadherin were investigated by Western Blot Results: Compared with human immortalized liver cell line L02, the expression of miR-30 lb in HCC cell lines were significantly increased. After being transfected with niiR-301b mimic, the expression of miR-301b was significantly improved than that of the control group. And the expression of rniR-301b was significantly decreased than that of the control group after being transfected with miR-301b inhibitor. The dual-luciferase reporter gene experiment showed that rniR-301b directly bound with Klf4's 3 TJTR region and decreased the expression of its protein, which were consistent with the software predicting results. The wound healing and Transwell experiment results showed that miR-301b inhibited the migration of hepatocellular carcinoma by downregulating Klf4. Further experiments showed that overexpression of rniR-301b significantly reduced the level of E-cadherin and increased the level of Nodherin, which were involved in the process of EMT. Conclusions: miR-301b was over-expressed in SK-Hep-1 and HCC-LM3 cells, and promoted the migration ability of HCC cells by inhibiting the expression of Klf4 gene, which may be involved in epithelial mesenchymal transformation progress of HCC cells.
- Publication
Progress in Modern Biomedicine, 2018, Vol 18, Issue 10, p1806
- ISSN
1673-6273
- Publication type
Article
- DOI
10.13241/j.cnki.pmb.2018.10.002