Phytotherapeutic potential of Pterospermum acerifolium L. in human cancer cell lines through mitochondrial mediated ROS generation

Abstract

Natural products have provided a new direction for the treatment of many human cancer including lung and pancreatic cancer. Phytotherapeutics provide a better alternative over conventional chemotherapeutics as these have less side effects. Pterospermum acerifolium commonly called as ‘Kanak champa’ have a wide range of application in Indian traditional medicinal system. P. acerifolium has many medicinal properties; however, the anticancer potential of P. acerifolium is not well studied. In the present study, we have demonstrated the anticancer effect of P. acerifolium bark extract in lung and pancreatic cancer cells. Objectives- (1) To evaluate the in vitro cytotoxic potential of Pterospermum acerifolium bark extract in lung and pancreatic cancer cell lines. (2) To investigate the role of Pterospermum acerifolium in ROS generation and mitochondrial mediated apoptosis in lung and pancreatic cancer cells. Methods- The anticancer potential of P. acerifolium bark extract on A549 lung and PANC-1 pancreatic cancer cell lines were determined by MTT assay, flow cytometry, fluorescence staining, cell cycle analysis, clonogenic assay, scratch invasion assay and quantative real time PCR study. Results and Conclusion- The cell viability by MTT assay showed that cytotoxicity was increased in a dose dependent manner in both lung and pancreatic cancer cell lines. Treatment with different concentrations of P. acerifolium bark extract showed clear visualization of change in cellular morphology, cell shrinkage, and cytoplasmic condensation in both A549 and PANC-1 cell lines. Scratch and Invasion assay showed that cell migration and invasion was significantly checked bark extract treated A549 and PANC-1 cells. Acridine orange/ethidium bromide and DAPI staining of P. acerifolium bark extract treated A549 and PANC-1 cells showed early and late apoptotic cells. Fluorescence and flow cytometry staining analysis showed that P. acerifolium bark extract reduces mitochondria membrane potential, induces ROS generation and increases the percentage of sub-G1 phase in both lung and pancreatic cancer cells confirming the apoptotic potential. In conclusion, we demonstrated that P. acerifolium bark extract showed cell cytotoxicity in lung and pancreatic cancer cells through modulating ROS generation, mitochondrial membrane potential and cell cycle checkpoints.