Exploration of CAM Complementary and Alternative Medicine Therapies against Colon Cancer an in Vitro Study using Colo 205 Cells

Abstract

The use of complementary and alternative medicine CAM for various ailments including cancer has been around for centuries To validate the ethno pharmacological claims against cancer four different CAM sources 1 Shoots of Triticum aestivum known as wheatgrass 2 Achyranthes aspera roots 3 Mushroom species Pleurotus ostreatus, Auricularia polytricha and Macrolepiota proce and 4 Homeopathic remedy Ruta graveolens, were investigated on colon cancer COLO 205 cells, as either individual extracts potency or as a cocktail mix newlineVarious in vitro approaches were adopted to investigate the most potent extract from each CAM source that possesses maximum cytotoxic, anti proliferative and apoptosis eliciting properties Trypan blue exclusion and MTT assay were used to assess cytotoxic potential Anti proliferation assays were carried out to evaluate the effect of CAM sources on COLO 205 cells, including clonogenic survival, wound scratch and trypan blue viability Induction of apoptosis was confirmed through various morphological methods by staining cells with various dyes such as, Hoechst, Acridine orange and Annexin V PI, while biochemical features were investigated by the qualitative and quantitative assessment of DNA fragmentation Based on the results of the above studies the extract potency showing maximal effect were further selected to gain an insight into the mechanism of action of cell death by performing RT PCR of apoptosis and cell cycle related genes, such as caspase 9, caspase 8, caspase 3, Bax, Bcl 2, p16, p21 and p27, followed by cell cycle analysis by flow cytometry newline newline