ADH1B Gene Promotes Gastric Cancer Tumorigenesis and its Cisplatin Resistance through the EMT Pathway.

BACKGROUND

Cisplatin resistance significantly affects the outcome of gastric cancer treatment. In this study, genes associated with cisplatin resistance were investigated and discussed.

METHODS

We analyzed the sequencing data of GSE14208 patients from the GEO database using differential and enrichment analyses. Gastric cancer cells with high ADH1B expression and low ADH1B expression were selected by qPCR and WB to construct ADH1B overexpress and silence cells. The optimal cisplatin concentrations for treatment were determined via CCK-8 detection and WB. Furthermore, we assessed drug resistance, cellular activity, and invasion and migration capacities using IC50, CCK-8 assays, Transwell, and migration tests. Apoptosis was evaluated using flow cytometry and EDU staining. The pathway influenced by ADH1B was examined through WB and immunofluorescence. The impact of gene expression on the tumorigenic potential of gastric cancer cells was assessed by analyzing tumor size, mass, and histology in HE-stained tumor sections and Ki67 and protein pathway immunohistochemistry in a mouse tumorigenesis model.

RESULTS AND DISCUSSION: This study revealed that ADH1B may exhibit a cancer-promoting effect, according to our database analysis, and is associated with drug resistance. Silencing ADH1B in AGS cells led to a reduced IC50, as well as decreased viability, invasion, and migration capabilities while increasing apoptosis rates. Conversely, overexpressing ADH1B in MKN-45 cells reversed these effects. Western blot and immunofluorescence results indicated that the expression levels of proteins involved in the EMT, PI3K, and MAPK pathways were altered following the silencing and overexpression of ADH1B. Moreover, silencing ADH1B not only reduced tumor volume and weight but also enhanced the tumor-reducing effects of cisplatin, as evidenced by changes in tumor structure; overexpression had the opposite effects. The alterations in pathway protein expression in tumor sections mirrored those observed in cells.

CONCLUSION

This study identified the gene ADH1B as a critical factor in the incidence and drug resistance of gastric cancer. It was demonstrated through cellular and tumorigenic assays that ADH1B promotes carcinogenesis and enhances cisplatin resistance in gastric cancer cells via the EMT signaling pathways.