Experimental Filtering Of MicroRNAs Targeting Pin1

Objective:Select mi RNAs which targeting regulation Pin1 for providing a new idea in the diagnosis and treatment of Hepatocelluar carcinomaMethods:1. To know the expression of Pin1 in HCC cell lines Hep G2, Huh7 and Hep3 B, we used Western blot assay and Q-PCR experiment to detect the Pin1 expression levels. 2. mi RNAs were selected by bioinformatic prediction sofeware and we conducted the following experiments, the double-luciferase reporter gene assay, Western blot test, and Q-PCR experiments to validation mi RNAs which could target on Pin1 3’UTR and regulate the expression of Pin1.Result:1. In HCC cell lines HepG2, Huh7 and Hep3 B, compared with normal liver cell line LO2, Western blot assay confirmed that Pin1 protein were overexpression, and QPCR experiments found that the expression of Pin1 m RNA in Hep G2 cells were lower than that of LO2 cells, while in Huh7 cells and Hep3 B cells, the relative expression of Pin1 m RNA were higher. 2.Four mi RNAs(mi R-140-5p, mi R-429, mi R-200 b, mi R-200c) were selected by bioinformatic prediction sofeware. The dual luciferase reporter gene assay and Western blot experiments showed mi R-140-5p obviously reduced luciferase activity and Pin1 protein in all screening mi RNAs.Conclusion:1.The expression of Pin1 protein in HCC cell lines were over-expression. 2. mi R-140-5p targeted regulation Pin1 gene.