Investigation of the effect of betulinic acid on expression of apoptosis-associated genes in kidney cancer cell line

Kidney cancer is the fourteenth most frequent cancer in the world. With its increasing incidence in the last few years, the renal cell carcinoma is the most common kidney cancer type. Apart from the environmental factors, increasing activities of expressions of genes and/or their pathways make cells become cancerous. AKT-1 and mTOR promote cell growth, proliferation and survival, participate in cellular homeostasis and have ability to suppress apoptosis in various ways. Herbal components are being used to reactivate apoptosis or suppress proliferation in cancer cells. One of the components is betulinic acid which is a lupane-type pentacyclic triterpenoid is derived from various plants. It is known that betulinic acid promotes apoptosis in cancer cells along with no toxic effect in normal cells. This effect makes betulinic acid a potential anti-cancer drug. In this study, kidney cancer cell line CAKİ-2 and normal cell line MRC-5 are used to research the apoptotic effect of betulinic acid in gene expression level. In order to identify the toxic effect of betulinic acid, WST-1 and to detect apoptosis Annexin-V were conducted. Expression analysis of AKT-1 and mTOR genes were conducted with Real-Time PCR to measure the effect of betulinic acid in gene level. As a result of this study, the apoptotic activity of betulinic acid on kidney cancer cell line was detected with Annexin-V. In gene expression analysis, a significant increase in mTOR gene expression levels was observed due to dose dependent, a small amount of decrement was observed in AKT-1 expression levels however the change is not statistically meaningful. Betulinic acid is a potential anti-cancer drug with its apoptotic effect on kidney cancer cell line and non-toxic effect on normal cell line and promising for future studies. Key Words: betülinic acid, kidney cancer, apoptosis, AKT-1, mTOR