Taxifolin binds with LXR (α & β) to attenuate DMBA-induced mammary carcinogenesis through mTOR/Maf-1/PTEN pathway.

Abstract

7,12-dimethylbenz(a)anthracene(DMBA), a PAH derivative initializes cascades of signaling events that alters a variety of enzymes responsible for lipid and glucose homeostasis resulting in enhanced availability and consumption of energy producing molecules for the development of carcinogenesis. 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoAR) is a key enzyme regulating the pathway of synthesis of cholesterol whereas liver-X-receptor (LXR) regulates lipid, carbohydrate metabolism in various malignancies including mammary carcinogenesis (MC). In this study Taxifolin (TAX), a potential flavanoid has been subjected to evaluate its anti-cancer potential on (MC). We designed to screen the molecular docking analysis of TAX on LXRα, LXRβ, HMG-CoAR, mTOR and PTEN using MAESTRO tool comparing with their reference ligands. MC was developed by the administration of DMBA in the air pouch (under the mammary fat pad) of the female Sprague-Dawley rats (55 days old). After 90 days of cancer induction, the chemotherapeutic potential of TAX was evaluated by administering TAX at different doses (10, 20 and 40 mg/kg b.w./day). Then western blot and RT-qPCR analysis were performed for determination of the protein and mRNA expressions respectively. The docking analysis revealed significant interaction with LXR (α&β), HMG-CoAR, mTOR and PTEN. The docking results were validated with the enzyme inhibition assay using HMG-CoAR (EC 1.1.1.34). TAX inhibited the HMG-CoAR activity with an IC50 value of 97.54 ± 2.5 nM whereas the reference molecule pavastatin revealed an IC50 value of 84.35 ± 1.2 nM. Moreover, TAX modulated the energy regulation on DMBA-induced MC in SD-rats by significantly restoring the cancer-induced alterations in body weight, tumor growth and lipid, lipoproteins, lipid metabolizing enzymes and glycolytic enzymes. TAX interacted with LXRs, HMG-CoAR, metabolic enzymes and restored the altered metabolism that accelerates uncontrolled cell proliferation in MC. Moreover, TAX also altered the mRNA and protein expressions of HMG-CoAR, LXR (α,β), Maf1, PTEN, phosphoinositide 3-kinase (PI3K), Akt, mTOR, fatty acid synthase (FASN) and acetyl-CoA carboxylase 1 (ACC1) in a dose dependent manner. These results validate the anti-cancer potential of TAX in DMBA-induced MC through LXR-mTOR/Maf1/PTEN axis.