Bhallatakadi modaka induces apoptosis of cancer cells via activating ROS/p-ERK pathway

Bhallatakadi modaka (BM) is an Ayurvedic formulation that has been traditionally used for various therapeutic purposes, but its anticancer effects and underlying molecular mechanisms remain insufficiently characterized.This study aimed to investigate the antitumor effects and underlying mechanisms of BM in cancer cell lines and a xenograft mouse model. In vitro assays, including MTT and clonogenicity assays, revealed that BM inhibited cancer cell growth and induced apoptosis in a dose-dependent manner. Flow cytometry, DAPI/PI staining, and Western blotting further confirmed apoptosis induction. Additionally, BM treatment reduced the cancer stem cell population, evidenced by a decrease in CD44+CD117+ cells and reduced sphere formation. Mechanistically, the formulation induced the generation of ROS, which activated the extracellular signal-regulated kinase (p-ERK) pathway, culminating in apoptosis. Increased expression of apoptosis markers such as cleaved caspase-3 and decreased total PARP1 further confirm the apoptotic effect. In vivo, treatment with BM in OVCAR-3 xenograft-bearing mice led to a significant reduction in tumor size compared to controls. Immunohistochemistry showed a marked decrease in Ki-67 expression, indicating suppressed tumor cell proliferation. Overall, the data suggest that BM may inhibit cancer cell growth through mechanisms involving ROS accumulation, p-ERK activation, and induction of apoptosis.