Sh-MARCH8 Inhibits Tumorigenesis via PI3K Pathway in Gastric Cancer

Abstract

Background/aims: To identify new treatment strategies for gastric cancer and to elucidate the mechanism underlying its pathophysiology, we transfected sh-MARCH8 into the human gastric cancer cell lines MKN-45 and AGS to investigate the roles of MARCH8 in gastric cancer.

Methods: We used genetic engineering to construct the sh-MARCH8 interference plasmid and transfected it into gastric cancer cells. Colony formation assays and cell viability measurements were performed to detect the viability and proliferation of cancer cells. Wound healing assays were performed to estimate the migration and proliferation rates of the cells. Cell invasion assays were used to estimate the invasive abilities of the cells. Cell apoptosis analysis was performed by using flowing cytometry. Western blot analysis was performed to estimate the expression levels of proteins. Statistical analysis was performed using the SPSS 18.0 software. Student's t-test was used to determine the significance of all pairwise comparisons of interest.

Results: We observed that the transfection of sh-MARCH8 inhibited the survival and proliferation of MKN-45 and AGS cells. The migration and invasion of the MKN-45 and AGS cells were significantly decreased, and apoptosis was induced in comparison with the control cells. These results were further confirmed by data showing that sh-MARCH8 increased the BAX/BCL2 ratio in MKN-45 and AGS cells. We also observed that sh-MARCH8 inactivated the PI3K and ß-catenin stat3 signaling pathways by changing protein expression levels or the phosphorylation of related proteins.

Conclusion: These data suggested that sh-March8 reduced viability and induced apoptosis of the MKN-45 and AGS cells through the PI3K and ß-catenin stat3 signaling pathways. Taken together, our data revealed that transfection of sh-MARCH8 into the MKN-45 and AGS gastric cancer cell lines inhibited their growth, and this approach may be useful as a novel strategy for gastric cancer therapy.

Keywords: Apoptosis; Gastric cancer; MARCH8; PI3K pathway; ß-catenin stat3 pathway.