Ethanol promotes migration and invasion in colorectal cancer cells via epithelial-mesenchymal transition

Abstract

Objective To determine the effects of ethanol on the cell proliferation, migration and invasion in human colorenctal cancer HT29 and SW480 cells and investigate the underlying mechanisms. Methods The effect of ethanol on the proliferation of HT29 and SW480 cells was detected by CCK-8 assay. After the IC50 value was obtained by the relevant analysis, and low concentration ethanol exposure models (1/10 IC50 value) were established. The cell cycle distribution was measured by flow cytometry, cell migration and invasion were detected by cell scratch test and Transwell chamber assay respectively, and the cell ROS production were detected using flow cytometry. The protein levels of epithelial-mesenchymal transition (EMT) related proteins, such as E-cadherin, N-cadherin and vimentin were detected by Western blotting. The production of acetaldehyde in the cells after ethanol treatment was detected with acetaldehyde detection kit. Results The IC50 values of ethanol to HT29 and SW480 cells were 436 and 424 mmol/L, respectively. With the increase of ethanol concentration, the proportion of cells at G1 stage was increased. Compared with the control group, the cell scratch test showed that the percentages of wound closure of HT29 and SW480 cells were increased significantly after ethanol treatment in each concentration group (P < 0.05). Transwell chamber test indicated that ethanol treatment significantly increased the number of HT29 and SW480 cells permeating without and with matrigel (P < 0.05). Further experiments showed that ethanol significantly enhanced the intracellular ROS level of HT29 and SW480 cells (P < 0.05). Western blotting displayed that ethanol decreased the expression level of E-cadherin, while increased those of N-cadherin and vimentin (P < 0.05). And as the concentration of ethanol increasing, the acetaldehyde content was elevated in the cells. Conclusion Ethanol may enhance the invasion and migration of colorectal cancer cells through ROS-mediated EMT pathway.

Keywords

• ethanol
• colorectal cancer
• migration
• invasion
• mesenchymal transition