Role of GSK-3β in hepatocellular carcinoma cell migration / 第二军医大学学报

ABSTRACT

Objective: To study the effects of GSK-3β on the migration ability of hepatocellular carcinoma cells. Methods: Inhibition tests with specific inhibitors of GSK-3 (LiCl and SB216763) and transfection with different plasmids(GSK-3β WT, GSK-3β S9A, and G1D5-6) were used to alter the activity of GSK-3β. The migration ability of SMMC-7221 and Hep3B cells was observed using wound healing assay, transwell assay and the microtubule- organizing center (MTOC) test before and after treatment. Immunocytochemistry was used to investigate the distribution of phospho-GSK-3β. Results: Wound healing assay showed that LiCl and SB216763 decreased the migration of SMMC-7721 cells by 36. 44% and 41. 78%, respectively; and decreased the migration of Hep3B cells by both 26. 66%. Transfection with GSK-3β WT increased the expression of GSK-3β and pGSK-3β, and the migration rate of SMMC-7721 cells decreased to 61. 27%. Transfection with GSK-3β S9A increased GSK-3β expression and had no noticeable influence on pGSK-3β expression, and the migration rate of SMMC-7721 cells decreased to 38. 61%. Transfection with GID5-6 had no noticeable influence on GSK-3β and increased pGSK-3β expression, and the migration rate decreased to 36. 49%. Immunocytochemical staining showed that LiCl blocked pGSK-3β accumulation on cell edge as that in normally cultured cells. MTOC test showed that the MTOC positive rate was above 60% in Hep3B cells before adding inhibitors of GSK-3β, and it decreased after exposure to the inhibitors. Conclusion: GSK-3β plays an important role in migration of hepatocellular carcinoma cells. Inhibitors of GSK-3β can affect the polarization and decrease the migration ability of hepatocellular carcinoma cells.