In vitro and in vivo evaluation of anticancer effects of KL21 polyherbal formulation and its toxicity potency

Inhibition of angiogenesis and induction of apoptosis are two of the latest strategies for cancer treatment. The discovery of novel angiogenic inhibitors with proapoptotic activities is therefore extremely crucial for opening up new avenues in this area. The present study is designed to provide a step-by-step methodology for evaluating a polyherbal formulation (KL21) for its potential as an anticancer agent that may later be developed into a pharmaceutical product. In this regard, as the first step, the total phenolic flavonoid content and antioxidant capacity of ethanol and methanol extracts of KL21 were determined with chromatographical and spectrophotometrical methods. DPPH and ABTS assays were utilized for the estimation of antioxidant capacity in both extracts. Additionally, MIC method was also carried out and the methanolic extract showed better antimicrobial activity. In cytotoxicity tests, MCF-7, HEK-293 and HeLa cell lines were used for calculating LD50 value. Extracts showed no genotoxic effect according to AMES test. Following the acute toxicity test which resulted in an LD50 value of ≥ 2000 mg/kg, a 28-day subacute toxicity test was conducted. At the end of this study, hematological and biochemical parameters were obtained and histopathological analyses were performed. Anticancer efficacy of KL21 was investigated on Ehrlich ascites tumor model. Tumor volumes were found to be decreased in post-implantation group of KL21 plyherbal formulation. The profiles of 84 genes involved in various apoptotic pathways were investigated via "Real Time Ready Human Apoptosis Panel". In another stage of the study, anti-angiogenic potencies of 900 mg/kg and 600 mg/kg dose groups were exhibited using the HET-CAM method. "Cyto Select Kit" was used in cell migration experiments and percent wound closure was calculated as 28%.