Screening methods for anti-kidney and adrenal cancer drug candidates

Abstract

An increased incidence of renal cell carcinoma (RCC) differs widely in various parts of the world, approximately 175,000 deaths because of kidney cancer in 2018. Early diagnosis while cancer is still localized rises the rate of survival and gave better chance to overcome the disease. CYP11A1 is a mitochondrial side-chain cleavage enzyme which are involved first step of steroid synthesis and associated as downstream regulation in various cancer types. Furthermore, the deficiency of CYP11A1 expression can cause adrenal insufficiency that lead to lethal when untreated. The purpose of our study is to develop an assay for screening natural product compounds that activate CYP11A1 by perform simultaneous analysis of cancer cell mobility, cytotoxicity and P450scc enzyme activity. We transfected CYP11A1 to the kidney cancer cell line (Caki-1) and overexpressed it to screen 1347 natural product compounds from Korea Chemical Bank. Using the untreated group and the group treated with aminoglutethimide, an inhibitor of CYP11A1, or mitomycin, an activator as controls, the compounds to activate CYP11A1 were selected. And their cancer mobility impair and cytotoxicity were evaluated at the same time. We identified 17 excellent anticancer candidates and will test IC50, Cholesterol and Pregnenolone level using multi reaction monitoring based LC-MS/MS approach. We observed that the cell migration and invasion were inhibited along with epithelial mesenchymal transition markers snail and vimentin were reduced by CYP11A1 overexpression. This finding might suggest new therapeutic target as well as anticancer drug candidates for kidney and adrenal cancers.