Effect And Mechanism Research Of Glibenclamide On The Growth Of Glioblastoma

Objective: The tumor is one of the four major diseases(heart disease,tumor,cerebrovascular disease,Alzheimer's disease),which serious harm to human health and formed by the abnormal cell differentiation,proliferation.Chemotherapy is the main method for the treatment of tumor and often cause many side effects,such as: low immune function,bone marrow suppression,and patients with drug resistance in the course of chemotherapy,which increased the difficulty of treatment.As a common type of brain tumor astrocytoma,which divided into four pathological grades by the WHO according to the histopathologic characteristics.And glioblastoma is the highest malignant glioma with aggressive,high incidence rate,postoperative and easy to relapse,high death rate and the clinical therapeutic effect unsatisfactory.Therefore,it is urgent to study the pathogenesis and explore new treatment methods.Methods: In this study,we investigated the effects of glibenclamide on U251 and U87 cells,and explored effect and mechanism of glibenclamide on the proliferation and growth of glioblastoma U251 and U87 cells.The application of CCK8,wound healing assay,transwell migration and invasion,hocheset staining,flow cytometry,pH probe,western blot and other experimental technology to dectet the effect and mechanism of glibenclamide on the proliferation and growth of U251 and U87 cells.The U251 and U87 cells of human glioma were cultured in vitro,and were randomly divided into experimental group and control group.U251 and U87 cells were treated with different concentrations of glibenclamide at 48 h in vitro,and the cell proliferation was measured by CCK8.The migration and invasion of cells were observed by transwell method and wound healing assay.Hocheset staining and flow cytometry were used to detect the changes of apoptosis.PH probe was used to observe the changes of intracellular pH.Western Blot was used to detect the expression levels of Kir4.1 and MCT1 in U251 and U87 cells.Results:CCK8 showed that in a certain concentration range,glibenclamide inhibited cell proliferation in a dose-dependent manner(P < 0.05);Wound healing assay showed glibenclamide inhibited cell migration(P < 0.05),and Transwell showed the number of cells in migration and invasion are positive with the drug concentration(P < 0.05);Hochest assay and flow cytometry showed characteristics of apoptosis appeared in the experimental group obviously(P < 0.05),and with the increase of glibenclamide concentration,apoptosis is more obvious(P < 0.05);Fluorescent probe showed that with the increase of drug concentration,the fluorescence intensity decreased(P < 0.05),suggesting that the higher drug concentration,intracellular pH decreased gradually(P < 0.05);Western blot results showed that the experimental group of the expression of Kir4.1 and MCT1 protein content decreased significantly(P < 0.05),and decreased with the increase of drug concentration(P < 0.05).Conclusion: 1 Glibenclamide inhibits proliferation of glioblastoma;2 Glibenclamide involved in the regulation of pH in the internal environment and plays a role in inhibiting proliferation by affecting the expression of Kir4.1 and MCT1.