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目的:研究14-3-3家族在乙型肝炎病毒(hepatitis B virus,HBV)复制的肝癌细胞中的表达。方法:运用q RT-PCR和Western blot分析14-3-3家族在HBV复制细胞系中的表达水平;q RT-PCR、Western blot检测HBV复制在Hep G2细胞中对14-3-3ζ表达的影响;Western blot检测HBV复制在小鼠肝组织中对14-3-3ζ表达的影响。结果:在Hep G2.2.15中,14-3-3η、γ、ζm RNA和蛋白的表达水平较Hep G2对照组明显增加(m RNA:t=16.5、27.86、17.23,P=0.000、0.000、0.000;蛋白:t=35.7、16.97、22.51,P=0.000、0.000、0.000);在Hep AD38(Tet-)中只有14-3-3ζm RNA和蛋白的表达水平较Hep AD38(Tet+)对照组明显增加(m RNA:t=14.27,P=0.000;蛋白:t=12.11,P=0.000);HBV复制能使Hep G2中14-3-3ζm RNA和蛋白的表达显著升高(m RNA:F=98.8,P=0.000;蛋白:F=118.3,P=0.000);HBV复制能使小鼠肝组织中14-3-3ζ蛋白的表达显著升高(F=109.4,P=0.000)。结论:HBV可以促进肝癌细胞内14-3-3η、γ、ζ的表达,这一发现为控制HBV感染的治疗策略提供新的靶点。
OBJECTIVE: To study the expression of 14-3-3 family in hepatocellular carcinoma cells replicated by hepatitis B virus (HBV). Methods: The expression of 14-3-3 family in HBV replication cell lines was analyzed by q RT-PCR and Western blot. The expression of 14-3-3ζ in Hep G2 cells was detected by q RT-PCR and Western blot Western blot was used to detect the effect of HBV replication on the expression of 14-3-3ζ in liver tissue of mice. Results: In Hep G2.2.15, the expression levels of 14-3-3η, γ, ζm RNA and protein were significantly increased compared with Hep G2 control group (m RNA: t = 16.5,27.86,17.23, P = 0.000,0.000,0.000 ; Protein: t = 35.7,16.97,22.51, P = 0.000,0.000,0.000); only the expression level of 14-3-3ζm RNA and protein in Hep AD38 (Tet-) was significantly higher than that in Hep AD38 (Tet +) control group (m RNA: t = 14.27, P = 0.000; protein: t = 12.11, P = 0.000); HBV replication increased the expression of 14-3-3ζm RNA and protein in Hep G2 (m RNA: F = 98.8 , P = 0.000; protein: F = 118.3, P = 0.000). HBV replication significantly increased the expression of 14-3-3ζ protein in the liver of mice (F = 109.4, P = 0.000). Conclusion: HBV can promote the expression of 14-3-3η, γ and ζ in hepatocellular carcinoma cells. This finding provides a new target for the treatment of HBV infection.