Identification of molecular pathways affected by treatment with heartwood water extract of Pterocarpus marsupium in MCF 7 cancer cell line

Abstract

The heartwood of Pterocarpus marsupium Roxb. has a history of traditional use in Ayurveda and traditional systems of medicines for the management of conditions such as diabetes and hyperlipidemia. The present study was conducted to identify the molecular pathways affected by a Heartwood Water Extract of P. marsupium (HWE). The effects of HWE on glucose uptake, as well as anti-oxidant and cytotoxic potential were studied in C2C12 and MCF-7 cell lines. An MCF-7 cell line synthesizing cholesterol and cholic acid, which generally occurs in liver cells, was treated with HWE and used to explicate the affected pathways through metabolomics.The study showed that treatment with HWE (100μg/mL) significantly increased antioxidant enzyme activity and increased glucose uptake activity by >3.0 fold (P<0.05). Treatment of MCF-7 cells with HWE (10mg/mL), also showed statistically significant apoptotic effects, with 78.2±2.4% (P<0.01) apoptotic cells at 24h. Metabolomics showed 36 metabolites to be differentially expressed to a significant degree after treatment with HWE. Amongst these, cholesterol synthesis intermediates, cholic acid derivatives, coenzyme A and sphingolipids were down-regulated (≥2 fold; P<0.05) in the HWE treated cell line. Histamine and carotene were found to be up-regulated by >2 fold. Nine pathways were found to be differentially regulated by HWE treatment, and cholic acid and sphingolipid pathways were identified as the primary targets of HWE.Decreased synthesis of cholic acid derivatives leads to less buffering and reduced absorption of lipids. The data suggests that reduced coenzyme A and increased histamine levels activated H+/K+ ATPase and that the resultant increase in energy consumption led the cells to apoptosis. The results also indicate that HWE regulates lipid and glucose homeostasis and suggest that HWE has a multi-faceted potential in the amelioration of hyperlipidemia and diabetes, as well as potential applications in cancer treatment.