Abstract LB-086: Genes targeted by drugs and curcumin in a breast carcinogenesis model

Abstract

Abstract Breast carcinogenesis is a multistage process that involves mutations and cellular phenotypic alterations attributed to exposure to exogenous environmental substances as well as endogenous agents as female hormones. Pamidronate (Pam), one of the nitrogen-containing bisphosphonates, is used in the treatment of bone metastases of breast cancer. It has recently been reported that it is also related to cell proliferation and apoptosis. 5-Fluorouracil (5-FU) is a chemotherapeutic agent for the treatment of a variety of solid cancers that arrest cell cycle and induce apoptosis in cancer cells. Curcumin (Cur) is an antioxidant known as a dietary natural yellow pigment derived from the rhizome of the herb Curcuma longa. The aim of this study was to evaluate genes that could be targeted by these drugs and curcumin in a breast carcinogenesis in vitro model induced by radiation and estrogen. Such model was developed with a normal immortalized breast epithelial cell line, MCF-10F that was exposed to low doses of high LET (linear energy transfer) alpha particles (150 keV/μm) of radiation, and cultured in presence of 17β-estradiol. This model consisted of the following cell lines: i) MCF-10F, ii) Alpha3, a malignant non-tumorigenic, iii) Alpha5, a tumorigenic one and iv) Tumor2, derived from Alpha5 injected into the nude mice. Previous results showed that Alpha5 and Tumor2 increased cell proliferation, presented anchorage independency, invasive capabilities and tumor formation in nude mice, as well as microsatellite instability and loss of heterozygosity in chromosomes 6, 8, 11 and 17 and mutations of c-Ha-ras and Rho-A among others. Pam, 5-FU and Cur were analyzed with MCF-10F by MTT indicating that the mean LD50 was 10, 2 and 30 μM after 48 hrs, respectively. Such substances inhibited migration and invasion in both Alpha5 and Tumor2 cell lines compared to the control MCF-10F and their counterparts, and also decreased c-Ha-ras, Rho-A, p53, Serpin-1 and Cav-1 gene expression in Alpha5 and significantly in Tumor2 cell lines by RT-qPCR. A significant apoptotic activity was observed by flow cytometry in Alpha5 and Tumor2 cell lines in comparison to control MCF-10F and their counterparts. These compounds had a direct antitumor and apoptotic effect on Bcl-xL and Bax by down-regulation of a transcription factor as NFĸB gene expression in Alpha5 and Tumor2 cell lines. It can be concluded that c-Ha-ras, RhoA, p53, Serpin-1 and Cav-1 genes were targeted by drugs and curcumin in a model transformed by low doses of alpha particles and estrogen. Such genes are involved in critical steps in breast carcinogenesis. Supported by FONDECYT #1120006 (GMC) and Ministry of Education (MINEDUC), Universidad de Tarapacá, Arica, Chile Citation Format: Gloria M. Calaf, Debasish Roy, Richard Ponce-Cusi. Genes targeted by drugs and curcumin in a breast carcinogenesis model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-086. doi:10.1158/1538-7445.AM2015-LB-086