MicroRNA-29a-3p Regulates Hepatocellular Carcinoma Progression Through NF-?B Pathway

Objectives To study the role of MicroRNA-29a-3p(miR-29a-3p)in the development of hepatocellular carcinoma(HCC),and to elucidate the signal transduction mechanism of its role.Methods Firstly,in order to investigate the function of miR-29a-3p in the development of hepatocellular carcinoma,Real-time fluorescence quantitative polynucleotide chain reaction(qRT-PCR)analysis technology was used to investigate the expression levels of miR-29a-3p in 21 samples of hepatocellular carcinoma caused by chronic alcohol abuse and 18 normal liver tissue samples.Secondly,in order to elucidate the signal transduction mechanism of miR-29a-3p in the development of hepatocellular carcinoma,use the target gene prediction software(Target Scan)and found that there is a binding site between miR-29a-3p and PTEN,in order to determine whether there is a functional relationship between miR-29a-3p and PTEN,use q RT-PCR to determine the expression level of PTEN m RNA in hepatocellular carcinoma and normal liver tissues;Construct a model of lentivirus transfected liver cancer cells and set up a blank control group,use q RT-PCR and Western Blot to detect the expression levels of PTEN m RNA and protein in the blank control group and the miR-29a-3p inhibitor transfected group;use the dual luciferase report analysis system to measure the activity of NF-?B in BC group,miR-con group,miR-29a-3p group and miR-29a-3p inhibitor group.Finally,in order to further clarify the effect of miR-29a-3p on the proliferation of liver cancer cells in vitro,the cell proliferation activity of the BC group,the miR-29a-3p transfected group,and the miR-29a-3p inhibitor transfected group was measured by MTT method,and the expression levels of VEGF and MMP-9 in BC group and miR-29a-3p inhibitor group were detected by q RT-PCR technology and Western Blot.Results 1 Compared with normal liver tissue,the expression level of miR-29a-3p in hepatocellular carcinoma caused by chronic alcohol abuse is down-regulated(P?0.05).2 The expression level of PTEN m RNA in hepatocellular carcinoma tissue is significantly higher than that in normal liver tissue(P?0.05),and after transfection with miR-29a-3p inhibitors inhibited miR-29a-3p in human BEL-7402 hepatoma cells,compared with the blank control group(BC),PTEN m RNA and protein expression were up-regulated(P?0.05).3 Western Blot analysis found that miR-29a-3p inhibitor reduced the phosphorylation level of p65(P?0.05),the double luciferase reporter gene assay results showed that miR-29a-3p can increase the activity of NF-?B signal transduction pathway factors,and miR-29a-3p inhibitors can inhibit the activity of NF-?B(P?0.05).4 The cell proliferation activity measured by MTT method showed that the cell growth rate was effectively reduced after transfection of miR-29a-3p mimics(P?0.05),after adding miR-29a-3p inhibitor,the growth rate of hepatoma cell line BEL-7402 significantly increased(P?0.05),Compared with the blank control group,miR-29a-3p inhibitors can significantly increase the m RNA and protein expression levels of VEGF and MMP-9(P?0.05).Conclusions 1 Mi R-29a-3p expression is down-regulated in hepatocellular carcinoma caused by chronic alcohol abuse.2 The down-regulation of miR-29a-3p increased PTEN expression and inhibited NF-?B signaling pathway activity.3 The up-regulation of miR-29a-3p expression can inhibit the proliferation of BEL-7402 hepatocellular carcinoma cell line in vitro.Figure 8;Table 2;Reference 200...