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Hydrogen sulfide (H 2 S) is a gaseous bacterial metabolite that reaches high levels in the large intestine. In the present study, the effect of H 2 S on the proliferation of normal and cancerous colon epithelial cells was investigated. An immortalized colon epithelial cell line (YAMC) and a panel of colon cancer cell lines (HT-29, SW1116, HCT116) were exposed to H 2 S at concentrations similar to those found in the human colon. H 2 S inhibited normal and cancerous colon epithelial cell proliferation as measured by MTT assay. The anti-mitogenic effect of H 2 S was accompanied by G 1 -phase cell cycle arrest and the induction of the cyclin-dependent kinase inhibitor p21 Cip . Moreover, exposure to H 2 S led to features characteristic of autophagy, including increased formation of LC3B +  autophagic vacuoles and acidic vesicular organelles as determined by immunofluorescence and acridine orange staining, respectively. Abolition of autophagy by RNA interference targeting Vps34 or Atg7 enhanced the anti-proliferative effect of H 2 S. Further mechanistic investigation revealed that H 2 S stimulated the phosphorylation of AMP-activated protein kinase (AMPK) and inhibited the phosphorylation of mammalian target of rapamycin (mTOR) and S6 kinase. Inhibition of AMPK significantly reversed H 2 S-induced autophagy and inhibition of cell proliferation. Collectively, we demonstrate that H 2 S inhibits colon epithelial cell proliferation and induces protective autophagy via the AMPK pathway.

Hydrogen Sulfide Lowers Proliferation and Induces Protective Autophagy in Colon Epithelial Cells