Effects of microRNA-145 on CD44 expression and hepatocellular carcinoma cells / 中华肝胆外科杂志

ABSTRACT

Objective To study the relationship of microRNA-145(miR-145) and CD44 in the hepatocellular carcinoma(HCC).Methods 13 clinical samples of HCC tissues and corresponding normal liver tissues were collected at department of General Surgery,Second Affiliated Hospital of Fujian Medical University from October 2015 to June 2016.The patients with positive expression of CD44 were studied.The expression levels of miR-145 and CD44 in HCC tissues and corresponding normal tissues were detected by real-time quantitative PCR.Western blot was performed to detect the expression of CD44 in HCC cells Hep G2 and SNU423.Biological information methods predicted whether miR-145 and CD44 have a targeted relationship.CD44 expression levels were detected after high expression of miR-145 in HCC cell line with positive expression of CD44.The vector and luciferase reporter genes were constructed to verify the interaction between miR-145 and CD44.The effects of miR-145 on proliferation in HCC cell lines with positive and negative CD44 expression were examined by tetramethylazoazole salt (MTT) assay.Results 8 of the 13 patients showed positive CD44 expression in HCC tissues.Compared with normal liver tissues,the relative expression of miR-145 in HCC tissues was significantly lower (0.998±0.010 vs.0.503±0.046,P＜0.05),and the relative expression of CD44 was higher (0.996±0.005 vs.1.878±0.108,P＜0.05).Bioinformatics suggested that miR-145 had a targeted relationship with CD44.High expression of miR-145 can significantly reduce the expression level of CD44 mRNA in HCC cell SNU423 (0.941±0.010 vs.0.515±0.021,P＜0.05) and down-regulate the expression of CD44 protein.Confirmed by luciferase reporter assay,CD44 is the target gene of miR-145.After transfection with miR-145 mimics,the proliferation of CD44 positive cell SNU423 was significantly inhibited (P＜0.05).Conclusions miR-145 can affect the proliferation of CD44 positive HCC cells by regulating the expression of CD44,which may be one of the pathogenesis of HCC.