Lithocholic bile acid inhibits lipogenesis and induces apoptosis in breast cancer cells
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It has amply been documented that mammary tumor cells may exhibit an increased lipogenesis. Biliary acids are currently recognized as signaling molecules in the intestine, in addition to their classical roles in the digestion and absorption of lipids. The aim of our study was to evaluate the impact of lithocholic acid (LCA) on the lipogenesis of breast cancer cells. The putative cytotoxic effects of LCA on these cells were also examined.
The effects of LCA on breast cancer-derived MCF-7 and MDA-MB-231 cells were studied using MTT viability assays, Annexin-FITC and Akt phosphorylation assays to evaluate anti-proliferative and pro-apoptotic properties, qRT-PCR and Western blotting assays to assess the expression of the bile acid receptor TGR5 and the estrogen receptor ERα, and genes and proteins involved in apoptosis (Bax, Bcl-2, p53) and lipogenesis (SREBP-1c, FASN, ACACA). Intracellular lipid droplets were visualized using Oil Red O staining.
We found that LCA induces TGR5 expression and exhibits anti-proliferative and pro-apoptotic effects in MCF-7 and MDA-MB-231 cells. Also, an increase in pro-apoptotic p53 protein expression and a decrease in anti-apoptotic Bcl-2 protein expression were observed after LCA treatment of MCF-7 cells. In addition, we found that LCA reduced Akt phosphorylation in MCF-7 cells, but not in MDA-MB-231 cells. We also noted that LCA reduced the expression of SREBP-1c, FASN and ACACA in both breast cancer-derived cell lines and that cells treated with LCA contained low numbers of lipid droplets compared to untreated control cells. Finally, a decrease in ERα expression was observed in MCF-7 cells treated with LCA.
Our data suggest a potential therapeutic role of lithocholic acid in breast cancer cells through a reversion of lipid metabolism deregulation.
KeywordsLithocholic acid MCF-7 cells MDA-MB-231 cells TGR5 activation Lipogenesis Breast cancer Gut microbiota
This study was funded by a grant from La Ligue Contre le Cancer de la Charente et de Loire Atlantique. Trang H. Luu was also funded by a research fellowship from the Vietnam Ministry of Education and Training and the Phu Tho Pharmacy College.
Compliance with ethical standards
Conflict of interest
The authors declare that there is no conflict of interest.
- 12.A.A. Goldberg, V.R. Richard, P. Kyryakov, S.D. Bourque, A. Beach, M.T. Burstein, A. Glebov, O. Koupaki, T. Boukh-Viner, C. Gregg, M. Juneau, A.M. English, D.Y. Thomas, V.I. Titorenko, Chemical genetic screen identifies lithocholic acid as an anti-aging compound that extends yeast chronological life span in a TOR-independent manner, by modulating housekeeping longevity assurance processes. Aging 2, 393–414 (2010)CrossRefPubMedPubMedCentralGoogle Scholar
- 13.V. Sepe, B. Renga, C. Festa, C. Finamore, D. Masullo, A. Carino, S. Cipriani, E. Distrutti, S. Fiorucci, A. Zampella, Investigation on bile acid receptor regulators. Discovery of cholanoic acid derivatives with dual G-protein coupled bile acid receptor 1 (GPBAR1) antagonistic and farnesoid X receptor (FXR) modulatory activity. Steroids 105, 59–67 (2015)CrossRefPubMedGoogle Scholar
- 16.H. Sato, A. Macchiarulo, C. Thomas, A. Gioiello, M. Une, A.F. Hofmann, R. Saladin, K. Schoonjans, R. Pellicciari, J. Auwerx, Novel potent and selective bile acid derivatives as TGR5 agonists: Biological screening, structure−activity relationships, and molecular modeling studies. J. Med. Chem. 51, 1831–1841 (2008)CrossRefPubMedGoogle Scholar
- 24.Rahimi A, Lee YY, Abdella H, Doerflinger M, Gangoda L, Srivastava R, Xiao K, Ekert PG, Puthalakath H. Role of p53 in cAMP/PKA pathway mediated apoptosis. Apoptosis 18, 1492–1499 (2013)Google Scholar
- 27.B.S. Tan, K.H. Tiong, H.L. Choo, F.F. Chung, L.W. Hii, S.H. Tan, I.K. Yap, S. Pani, N.T. Khor, S.F. Wong, R. Rosli, S.K. Cheong, C.O. Leong, Mutant p53-R273H mediates cancer cell survival and anoikis resistance through AKT-dependent suppression of BCL2-modifying factor (BMF). Cell Death Dis. 6, e1826 (2015)CrossRefPubMedPubMedCentralGoogle Scholar
- 28.C.R. Loehberg, P.L. Strissel, R. Dittrich, R. Strick, J. Dittmer, A. Dittmer, B. Fabry, W.A. Kalender, T. Koch, D.L. Wachter, N. Groh, A. Polier, I. Brandt, L. Lotz, I. Hoffmann, F. Koppitz, S. Oeser, A. Mueller, P.A. Fasching, M.P. Lux, M.W. Beckmann, M.G. Schrauder, Akt and p53 are potential mediators of reduced mammary tumor growth by cloroquine and the mTOR inhibitor RAD001. Biochem. Pharmacol. 83, 480–488 (2012)CrossRefPubMedGoogle Scholar
- 30.M. Agostini, L.Y. Almeida, D.C. Bastos, R.M. Ortega, F.S. Moreira, F. Seguin, K.G. Zecchin, H.F. Raposo, H.C. Oliveira, N.D. Amoêdo, T. Salo, R.D. Coletta, E. Graner, The fatty acid synthase inhibitor orlistat reduces the growth and metastasis of orthotopic tongue oral squamous cell carcinomas. Mol. Cancer Ther. 13, 585–595 (2014)CrossRefPubMedGoogle Scholar
- 32.R. Singh, V. Yadav, S. Kumar, N. Saini, MicroRNA-195 inhibits proliferation, invasion and metastasis in breast cancer cells by targeting FASN, HMGCR, ACACA and CYP27B1. Sci. Rep. 5, 1–15 (2015)Google Scholar
- 34.T. Yamamoto, H. Shimano, N. Inoue, Y. Nakagawa, T. Matsuzaka, A. Takahashi, N. Yahagi, H. Sone, H. Suzuki, H. Toyoshima, N. Yamada, Protein kinase A suppresses sterol regulatory element-binding protein-1C expression via phosphorylation of liver X receptor in the liver. J. Biol. Chem. 282, 11687–11695 (2007)CrossRefPubMedGoogle Scholar
- 37.L. Tirinato, C. Liberale, S. Di Franco, P. Candeloro, A. Benfante, R. La Rocca, L. Potze, R. Marotta, R. Ruffilli, V.P. Rajamanickam, M. Malerba, F. De Angelis, A. Falqui, E. Carbone, M. Todaro, J.P. Medema, G. Stassi, E. Di Fabrizio, Lipid droplets: a new player in colorectal cancer stem cells unveiled by spectroscopic imaging. Stem Cells 33, 35–44 (2015)CrossRefPubMedGoogle Scholar
- 38.H. Abramczyk, J. Surmacki, M. Kopec, A.K. Olejnik, K. Lubecka-Pietruszewska, K. Fabianowska-Majewska, The role of lipid droplets and adipocytes in cancer. Raman imaging of cell cultures: MCF10A, MCF7, and MDA-MB-231 compared to adipocytes in cancerous human breast tissue. Analyst 140, 2224–2235 (2015)CrossRefPubMedGoogle Scholar