拉米夫定抑制HBV相关性肝癌HepG2.2.15细胞病毒复制、细胞增殖的实验研究

Inhibitory effect of lamivudine on replication and cell proliferation of HepG2.2.15 cells associated with HBV

目的:探讨拉米夫定抑制人乙肝病毒(HBV)相关性肝癌HepG2.2.15细胞病毒复制、细胞增殖的实验效果及其作用机制.方法:取对数生长期的HBV相关性肝癌HepG2.2.15细胞,分为实验组 A (拉米夫定100μg/ml)、实验组B (拉米夫定200μg/ml)、实验组C (拉米夫定300μg/ml)及空白组,在培养24h、 48h后分别检测HBV-DNA拷贝数、培养48h后的HBsAg、 HBeAg表达、 HepG2.2.15细胞凋亡率、细胞及上清液中的基质金属蛋白酶9 (MMP-9)、 p53蛋白表达水平.结果:在培养24h、 48h,空白组的HBV-DNA拷贝数均显著的高于A、 B、 C三组,差异具有统计学意义(P<0.05);A、 B、 C三组HBV-DNA拷贝数组间呈降低趋势,组间两两比较差异均具有统计学意义(P<0.001);在培养48h,空白组的HBsAg、 HBeAg水平均显著的高于A、 B、 C三组,差异具有统计学意义(P<0.001);A、 B、 C三组HBsAg、 HBeAg水平组间呈降低趋势,组间两两比较差异均具有统计学意义(P<0.001);在培养48h,空白组的HepG2.2.15细胞凋亡率均显著的低于A、 B、 C三组,差异具有统计学意义(P<0.001);A、 B、 C三组HepG2.2.15细胞凋亡率组间呈升高趋势,组间两两比较差异均具有统计学意义(P<0. 001);在培养48h,空白组的细胞及上清液中MMP-9、 p53蛋白表达均显著的高于A、 B、 C三组,差异具有统计学意义(P<0.001);A、 B、 C三组细胞及上清液中MMP-9、 p53蛋白表达组间呈降低趋势,组间两两比较差异均具有统计学意义(P<0.001).结论:拉米夫定能显著抑制HBV相关性肝癌HepG2.2.15细胞病毒复制,降低HBsAg、HBeAg、 MMP-9表达和增强p53蛋白的表达,对于促进肝癌HepG2.2.15细胞凋亡具有显著作用.

Objective:To explore the experimental effect and mechanism of lamivudine inhibiting viral replication and cell proliferation in human HBV-related liver cancer HepG2.2.15 cells.Methods:HBV-related liver cancer HepG2.2.15 cells in logarithmic growth phase were divided into experimental group A(lamivudine 100μg/nil),experimental group B(lamivudine 200μg/nil),experimental group C(lamiv)Set 300μg/ml)and blank group,HBV-DNA copy number,HBsAg,HBeAg expression,HepG2.2.15 apoptosis rate,matrix metalloproteinase-9(MMP-9)and p53 protein in cells and supernatant were detected after 24h and 48h.Results:At 24h and 48h,the HBV-DNA copy number of the blank group was significantly higher than that of A,B and C groups,and the difference was statistically significant(P<0.001).The A,B,and C groups showed a decreasing trend among the HBV-DNA copy arrays,and the differences between the two groups were statistically significant(P<0.001).After 48 hours of culture,the levels of HBsAg and HBeAg in the blank group were significantly higher than those in the A,B and C groups,and the difference was statistically significant(P<0.001).The levels of HBsAg and HBeAg in the three groups A,B and C showed a decreasing trend,and the differences between the two groups were statistically significant(P<0.001).The apoptosis rate of HepG2.2.15 cells in blank group was significantly lower than that in A,B and C groups at 48h,the difference was statistically significant(P<0.001).The apoptosis rate of HepG2.2.15 cells in group A,B and C showed an increasing trend,and the differenee between the two groups was statistically significant(P<0.001).The expressions of MMP-9 and p53 protein in the cells and supernatant of the blank group were significantly higher than those in A,B and C groups at 48 h,the differenee was statistically significant(P<0.001).The expression of MMP-9 and p53 protein in the cells of A,B and C groups showed a decreasing trend,and the difference between the two groups was statistically significant(P<0.001).Con elusion:Lamivudine significantly inhibited the viral replication of HBV-related liver cancer HepG2.2.15 cells,reduced the expression of HBsAg,HBeAg,MMP-9 and enhanced the expression of p53 protein,which has a significant effect on promoting apoptosis of HepG2.2.15 cells.