戈米辛G抑制HepG2细胞生长的机制研究

Mechanism of Gomisin G inhibiting the growth of HepG2 cells

目的探讨戈米辛G对HepG2细胞生长的抑制作用。方法将WRL68细胞和HepG2细胞分别分为3组:WRL68-1组(DMSO 10μL)、WRL68-2组(5μmol·L^(-1)Gomisin G 10μL)、WRL68-3组(10μmol·L^(-1)Gomisin G 10μL)和HepG2-1组(DMSO 10μL)、HepG2-2组(5μmol·L^(-1)Gomisin G 10μL)、HepG2-3组(10μmol·L^(-1)Gomisin G 10μL)。用MTT法检测WRL68细胞和HepG2细胞的增殖情况,并计算戈米辛G对于HepG2细胞的IC_(50);用流式细胞术检测HepG2细胞的凋亡情况;用蛋白质印迹法检测HepG2细胞Cyclin D1、bcl-2蛋白、Akt^(p-Ser473)和Akt蛋白的表达情况。结果WRL68-1组、WRL68-2组、WRL68-3组细胞24 h时的增殖率分别为(45.00±0.12)%,(42.55±4.85)%和(44.02±1.52)%,组间差异无统计学意义(P>0.05);HepG2-1组、HepG2-2组、HepG2-3组细胞24 h时的增殖率分别为(55.01±8.97)%,(48.82±6.14)%和(17.73±2.50)%,组间差异有统计学意义(P<0.05)。戈米辛G对于HepG2细胞作用24 h的IC_(50)约为8.93μmol·L^(-1)。HepG2-1组、HepG2-2组和HepG2-3组的细胞凋亡率分别为(0.64±0.14)%,(3.58±0.67)%和(8.67±1.11)%;这3组的Akt^(p-Ser473)灰度分别为1.02±0.27,0.89±0.16和0.32±0.06。结论戈米辛G能够通过抑制Akt^(p-Ser473)抑制HepG2细胞的生长。

Objective To investigate the inhibitory effect of Gomisin G on the growth of HepG2 cells.Methods WRL68 and HepG2 cells were divided into three groups:WRL68-1 group(DMSO 10μL),WRL68-2 group(5μmol·L^(-1) Gomisin G 10μL),WRL68-3 group(10μmol·L^(-1),Gomisin G,10μL)and HepG2-1 group(DMSO 10μL),HepG2-2 group(5μmol·L^(-1) Gomisin G 10μL),HepG2-3 group(10μmol·L^(-1),Gomisin G,10μL).MTT assay was used to detect the proliferation of WRL68 cells and HepG2 cells,and the IC_(50) of Gomisin G to HepG2 cells was calculated;the apoptosis of HepG2 cells was detected by flow cytometry;the expressions of Cyclin D1,bcl-2,Akt^(p-Ser473) and Akt in HepG2 cells were detected by Western blot.Results The proliferation rates of WRL68-1 group,WRL68-2 group and WRL68-3 group at 24 h were(45.00±0.12)%,(42.55±4.85)%and(44.02±1.52)%,respectively.The proliferation rates of HepG2-1 group,HepG2-2 group and HepG2-3 group at 24 h were(55.01±8.97)%,(48.82±6.14)%and(17.73±2.50)%,respectively and there was no significant difference between the groups(P>0.05).The IC_(50) of Gomisin G on HepG2 cells for 24 h was about 8.93μmol·L^(-1).The levels of apoptosis in HepG2-1 group,HepG2-2 group and HepG2-3 group were(0.64±0.14)%,(3.58±0.67)%and(8.67±1.11)%,respectively;the gray levels of Akt^(p-Ser473) in the three groups were 1.02±0.27,0.89±0.16 and 0.32±0.06,respectively.Conclusion Gomisin G can inhibit the growth of HepG2 cells by inhibiting Akt^(p-Ser473).