The effect of intermittent hypoxia and fecal microbiota of OSAS on genes associated with colorectal cancer.

ABSTRACT

PURPOSE: Colorectal cancer (CRC) is one of the common causes of cancer death worldwide. Obstructive sleep apnea syndrome (OSAS), sharing many risk factors in common with CRC, is prevalent among CRC patients. OSAS may promote the CRC development independently but the mechanism is still unknown. Intermittent hypoxia (IH) is one of the characteristics of OSAS, and hypoxia may influence the genes associated with CRC. Intestinal microbiota plays important role in CRC carcinogenesis, and OSAS patients have been shown to have intestinal microbiota dysbiosis. We hypothesized that IH and intestinal microbiota dysbiosis may be involved for CRC in patients with OSAS. METHODS: We established precancerous cell models of CRC with Immorto-Min colonic epithelial (IMCE) cells. First, the cells were exposed to IH in a special chamber for 4 h, 8 h, and 12 h. Feces from 6 patients with OSAS and 6 healthy controls were collected and made into sterile fecal fluid for incubation with IMCE cells for 12 h. The cells were then exposed to IH for 4 h, 8 h, and 12 h. After IH exposure, the expressions of genes and inflammation cytokines associated with CRC, such as ß-catenin, STAT3, HIF-1α, IL-6, TNF-α, c-myc, and cyclinD1, were tested. RESULTS: IH activated the expression of HIF-1α and STAT3 both in mRNA and protein level (HIF-1α P = 0.015 for mRNA level, P = 0.027 for protein level; STAT3 P = 0.023 for mRNA level, P = 0.023 for protein level), and promoted p-STAT3 shifting to the nucleus (P = 0.023). The mRNA of ß-catenin (P = 0.022) and cyclinD1 (P = 0.023) was elevated, but there was no change for the ß-catenin protein in the nucleus. Gut microbiota of OSAS patients promoted the expression of STAT3 (protein level 0 h P = 0.037; 4 h P = 0.046; 8 h P = 0.049; 12 h P = 0.037), promoted p-STAT3 (4 h P = 0.049; 8 h P = 0.046; 12 h P = 0.046) shifting to the nucleus, and also elevated the expression of IL-6 and TNF-α in mRNA level at 4 h (IL-6 P = 0.037, TNF-α P = 0.037) and 8 h (IL-6 P = 0.037, TNF-α P = 0.037). The protein of ß-catenin in the nucleus was not affected by IH and gut microbiota from OSAS. CONCLUSIONS: Our study demonstrated that IH and gut microbiota of patients with OSAS activated HIF-1α expression and STAT3 pathway in IMCE cells, with no influence on ß-catenin pathway, which suggested that IH, STAT3 pathway, chronic inflammation, and intestinal microbiota dysbiosis may be involved in CRC carcinogenesis correlated with OSAS These findings must be interpreted cautiously and further research is necessary to clarify the causative steps in CRC development.