The Effects Of Glucose Regulated Protein 78kD On Hepatoma Cell Proliferation, Autophagy And Angiogenesis In Vitro

Objective: To investigate the effects of suppressing GRP78 expression by si RNA on proliferation, autophagy, and angiogenesis of hepatoma cell, and providing new targets and new ideas for exploring effective drugs for antitumor and anti tumor angiogenesis.Methods: Four si RNA sequences specifically targeting GRP78(named as si GRP78-1,si GRP78-2,si GRP78-3,si GRP78-4) were transfected into hepatoma cell with Lipofectamine 3000.The best one of silent GRP78 was si GRP78-1. There were three groups in this study including blank control group, negative group, and si GRP78 group. The cell proliferation was assessed by MTT. The cell cycle was examined by flow cytometry. The expressions of GRP78, LC3-Ⅱ, AKT, P-AKT, and cyclin D1 were detected by Western blot. The ability of HUEVC migration and proliferation were surveyed by wound healing test and plate colony formation test.Results:The expression of GRP78 can be surpressed specifically by GRP78 si RNA; MTT results showed that the cell proliferation of MHCC97 H decreased in si GRP78 group(P<0.001);And the flow cytometry results showed that the G0/G1 phase cells of huh7 and SMMC7721 increased after being transfected with si GRP78(P<0.001,P<0.05); The expressions of LC3-Ⅱ, P-AKT, and cyclin D1 were decreased significantly in si GRP78 group; Wound healing test and plate colony formation test results showed that the ability of HUEVC migration and plate colony formation was inhibited in si GRP78 group.Conclusion: GRP78 si RNA surpresses specifically the expression of GRP78, inhibites cell proliferation, induces cell cycle arrest of huh7 and SMMC7721; And the mechanism may be that si GRP78 influences the PI3K/AKT signal pathway,and decreases the expressions of Cyclin D1; The suppression of GRP78 expression of huh7 inhibites the autophagy and the angiogenesis in the tumor micrornvironment.