A Study On Metformin Enhences Sorafenib’s Inhibition Of Hepatocellular Carcinoma By Targeting MicroRNA-200a

Objective Hepatocellular carcinoma is a malignant tumor with poor prognosis.Drug resistance of hepatocellular carcinoma is an important factor affecting the prognosis of patients.The pace of how to increase the sensitivity of liver cancer drugs has never stopped.In order to study that metformin can enhance the inhibition of the progression of hepatocellular carcinoma by Micro RNA-200 a and the therapeutic effect of sorafenib,and to explore the possible related mechanisms,Provide a theoretical basis for metformin to increase the level of Micro RNA-200 a to enhance the potential clinical application value of sorafenib in the efficacy of hepatocellular carcinoma.Methods To construct human liver cancer cell miR200 a stably transfected cell line:miR200 a overexpression,control,miR200 a knockdown group.RT-q-PCR method was used to detect the expression of miR200 a in each transfected cell line,MTT method was used to detect the effect of miR200 a on cell viability of each transfection group,and Western Blot analyzed the expression of each target protein;Transwell method was used to detect the influence of miR200 a on the migration and invasion ability of cells in each transfection group.Sorafenib was added to the transfected cell line,and MTT,RT-q-PCR and Western Blot were retested to verify that miR200 a regulates the viability of sorafenib on liver cancer cells and changes in the expression of corresponding genes and proteins;Add metformin to the transfected cell lines,retest MTT,RT-q-PCR and Western Blot to verify that metformin regulates the expression of miR200 a in each transfected group and its effect on the viability and corresponding protein expression of liver cancer cells;Finally,metformin and sorafenib were combined in the cells of each transfection group,and MTT,RT-q-PCR and Western Blot were retested again.To verify whether metformin strengthens sorafenib to regulate the expression of miR200 a in cells of each transfection group and its effect on the viability of liver cancer cells and corresponding protein expression.54 SPF BALB/c-nu male mice were randomly divided into 3 groups(transfected cell control group,transfected cell combine sorafenib group,transfected cell combine metformin with sorafenib group),There are 18 animals in each group,3 groups in each group,6 animals in each group.Each group of nude mice was subcutaneously planted with miR200 a overexpression,control,and miR200 a knockdown cells.After tumor formation,the placebo(normal saline)group and sorafenib group and metformin + Sorafenib combination group were injected daily to each group of nude mice;For 14 consecutive days,the nude mice were sacrificed,the tumor was taken out,and the tumor volume was calculated.RT-q-PCR,Western Blot,immunohistochemistry,immunofluorescence,etc.verify the changes in the transcription and translation levels of tumor-related genes.Results Up-regulation of miR200 a can not only inhibit the cell viability and migration and invasion ability of human liver cancer cells;it can also reduce the expression of SIRT1;Up-regulating the level of miR200 a significantly reduces the tumor volume in nude mice;up-regulating miR200 a can enhance the efficacy of sorafenib;metformin can up-regulate the level of miR200 a,further enhancing the drug effect of sorafenib.Conclusion Metformin can up-regulate the expression level of mi R200 a in liver cancer cells,negatively regulate the proliferation,cloning,migration and invasion of human liver cancer cells,and increase the efficacy of sorafenib.This effect may be achieved by regulating the expression of SIRT1.