Microbial metabolite deoxycholic acid inhibits noncancerous NCM460 human colon cell proliferation: an inverse correlation between Bmal1:Clock gene expression and cell apoptosis

Authors: Zeng, Huawei; Safratowich, Bryan; LIU, ZHENHUA - University Of Massachusetts; Briske Anderson, Mary

Submitted to: Archives of Biochemistry and Biophysics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/5/2025
Publication Date: 12/5/2025
Citation: Zeng, H., Safratowich, B.D., Liu, Z., Briske Anderson, M.J. 2026. Microbial metabolite deoxycholic acid inhibits noncancerous NCM460 human colon cell proliferation: an inverse correlation between Bmal1:Clock gene expression and cell apoptosis. Archives of Biochemistry and Biophysics. 776. Artlcle 110694. https://doi.org/10.1016/j.abb.2025.110694.
DOI: https://doi.org/10.1016/j.abb.2025.110694

Interpretive Summary: Colorectal cancer (CRC) is one of the most common cancers diagnosed in both men and women worldwide. Colon carcinogenesis involves several critical factors such as DNA damages and inflammation, and excess dietary fat intake is tightly associated with CRC risk. However, the underlying molecular mechanism between high-fat diet and CRC development remains to be determined. In this study, we demonstrated a potential cellular mutation process caused by second bile acids (because of high-fat meal consumption) in the colon. These data provide further insights into nutrition and obesity-related chronic diseases in the colon, and will be useful for scientists who are interested in diet and gut health.

Technical Abstract: High fat diets increase colonic deoxycholic acid (DCA) concentrations, which may induce apoptosis and enrich a compensatory DCA-resistant mutant colon cell subpopulation. However, the molecular mechanisms of DCA-induced apoptosis in normal colon cells remain elusive. Because circadian Bmal1 and Clock genes are key regulators for cell proliferation, we hypothesized that DCA-induced apoptosis in the colon is driven by Bmal1::Clock gene regulation. In this study, we determined the impact of DCA on the noncancerous human NCM460 colon cells. Compared to the untreated (control) cells, the treatment with DCA at 0.3- and 0.4-mM inhibited cell proliferation (via increasing cell cycle arrest and apoptosis) by 19% and 29%, respectively. As the Bmal1::Clock complex and the Wnt pathway are intimately interlinked with apoptotic processes, we identified that 42 (out of 84) genes were differentially expressed in the Wnt signaling pathway; and there was a decrease (= 47%) in Bmal1, Clock and Wee1 protein levels but an increase (= 172%) in Rev-Erb alpha protein levels in DCA-treated NCM460 cells compared to the control cells. At the clinical level, the mRNA levels of Bmal1 and Rev-Erb alpha (but not Clock) were decreased by at least 15% while Wee1 was increased by 38% in cancerous colon tissues compared to normal ones. Collectively, DCA increases cell cycle arrest and apoptosis accompanied with a drop of Bmal1::Clock gene expression and altered Wnt signaling pathway. The Bmal1::Clock regulatory network is relatively normal in the DCA-treated noncancerous colon cells but not in colon cancer tissues.