| Aim：To elucidate how and the mechanism by which nutrient-deprivationaffect invasions of hepatocellular carcinoma (HCC) cell lines.Methods：HCC cell lines HepG2and BEL-7402are cultured in Hank’s balancedsalt solution which serves as a way of nutrient deprivation. The effectsof nutrient deprivation on autophagic activities, invasions andexpressions of epithelial and mesenchymal markers of HCC cells areobserved. Then Atg3targeting siRNA (siRNA-Atg3) are constructed and usedto inhibit the autophagic activities of HCC cells, and the changes of HCCcells’ autophagic activities, invasion and expressions of epithelial andmesenchymal markers are recorded seperately before and after HCCcells’being transfected with siRNA-Atg3. The roles which autophagy andepithelial-mesenchymal transition play in nutrient deprivation-inducedinvasions of HCC cells are discussed.Results： Nutrient deprivation markly increase the numbers of LC3fluorescent puncta in both HepG2and BEL7402cell lines, which arefolloweded by increases in amount of LC3-Ⅱ protein and decreases inamount of p62protein. Nutrient deprivation also increases the numbersof HCC cells that invade through the membranes of Transwell chambers(52632±5238，p<0.05；45223±3370，p<0.05), and downregulates theexpression of epithelial marker CK18protein and upregulates theexpression of mesenchymal marker fibronectin and MMP9protein. Thetransfection of siRNA-Atg3can reverse HCC cells’ nutrientdeprivation-induced increases in amount of LC3-Ⅱ and decreases in amount of p62. The changes of expressions of epithelial marker CK18andmesenchymal markers fibronectin, MMP9, as well as the increase in numbersof HCC cells that invade through the membranes of Transwell chambers(21746±2619，p<0.05；20014±882，p<0.05) during nutrient deprivationare also inhibited by siRNA-Atg3.Conclusion：Autophagy-dependent epithelial-mesenchymal transition is themain mechanism by which nutrient deprivation promotes invasions of HCCcells. |
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