Anticancer effects of a traditional Thai herbal recipe Benja Amarit extracts against human hepatocellular carcinoma and colon cancer cell by targeting apoptosis pathways

Abstract

Ethnopharmacological relevance: A preparation of Benja Amarit (BJA) has been effectively used in folk medicine to treat diseases related to the liver and colon and forms of cancer for hundreds of years in Thailand. However, there has not been any research on BJA with regard to its anticancer activity against human hepatocellular carcinoma and colon cancer cells.

Aim of the study: This study was to obtain the scientific supports for the traditional usage in anticancer potential of BJA extracts on hepatocellular carcinoma and colon cancer.

Materials and methods: The cytotoxic effects were determined using MTT assay. Apoptosis was quantitated by annexin V-FITC/PI staining. Caspases activities were measured by using specific substrates and colorimetric analysis. The protein expressions were determined by Western blot analysis. Reactive oxygen species (ROS) generation, mitochondrial transmembrane potential, and calcium ion levels were measured by specific fluorescence probes and flow cytometry. The chick embryo chorioallantoic membrane model has been used to study the in vivo anticancer activity. The phytochemical identification was performed by GC-MS and LC-MS.

Results: Notably, 95% (BJA-95) and 50% (BJA-50) ethanolic extract of BJA inhibited hepatocellular carcinoma and colon cancer cell viability in a dose-dependent manner. While, the water extract of BJA (BJA-W) was not found to be toxic to both kinds of cancer cell lines. BJA extract induced both the extrinsic and intrinsic or mitochondria-mediated apoptosis pathways. Moreover, BJA-95 caused ROS generation and endoplasmic reticulum stress-mediated apoptosis. The extract exhibited the growth inhibitory effects on cancer cells in vivo. Phytochemical analysis revealed that the major active compounds were piperine, xanthotoxol and dihydrogambogic acid.

Conclusion: This study is the first to demonstrate anticancer efficiency of BJA extracts on human cancer cells. We consider BJA extract to be a potentially alternative cancer treatment and to be a promising candidate in the future development of antitumor agents.

Keywords: Apoptosis; BJA; Benja Amarit; Colon cancer; Hepatocellular carcinoma.