Liposomes encapsulated chloroform fraction of Cassia fistula L. bark enhances anticancer efficacy in ehrlich ascites carcinoma mice model

Cancer remains one of the eminent causes of mortality and a major health concern profoundly influencing life expectancy worldwide. Cassia fistula L. commonly known as ‘Golden shower' finds its application in the Ayurveda and Chinese traditional medicines. The phytoconstituents from the bark powder were obtained using organic solvents in the increasing order of polarity viz. CaMH (hexane fraction), CaMC (chloroform fraction), CaME (ethyl acetate fraction) and CaMM (methanol fraction). These bark fractions were tested for their cytotoxic potential in HeLa (cervical cancer), MCF-7 (breast cancer), LN-18 (brain glioblastoma), MG-63 (osteosarcoma), A431 (skin cancer), MIA PaCa-2 (pancreatic cancer) and normal cell line L929 (human fibroblast cells) using MTT assay. Among all the tested fractions, CaMC fraction shows highest selective cytotoxicity against HeLa cells with GI 50 value of 41.53 µg/mL. Silica gel column chromatography of CaMC fraction yielded Cf-1 fraction spectroscopically deciphered as Piperine. It showed significant cytotoxicity against HeLa, MG-63 and MCF-7 cell line with GI 50 of 0.10 mM, 0.37mM, 0.56 mM respectively. CaMC and Cf-1 fractions treated HeLa cells showed significant manifestations of apoptosis in morphological and flow cytometric investigations. These induced cell cycle arrest at G 0 /G 1 phase in concentration-dependent manner. Molecular mechanistic studies of CaMC and Cf-1 fraction depicted upregulation of apoptotic markers including p53, caspase 3 and 9, and downregulation of Bcl-2. Furthermore, the in-vivo studies evaluated in Ehrlich Ascites Carcinoma-bearing Swiss albino mice showed that liposomal encapsulated CaMC fraction ( CaMC-LP ) resulted in 82.23% tumor growth inhibition in contrast to CaMC fraction with 60.81% tumor inhibition, comparable to 86.39% inhibition by 5-Fluorouracil. The use of liposomal encapsulation enhanced the anticancer efficacy in vitro, and in vivo showing significant tumor inhibition in EAC- bearing mice without inducing toxicity. These findings suggest that C. fistula L. bark constituents have cancer chemopreventive and chemotherapeutic potential which can further be exploited for the benefit of mankind.