【摘要】 目的研究乙型肝炎病毒羧基末端155位截短的中表面蛋白(MHBst155)对肝癌细胞生长、增殖的影响。方法以Hep G2细胞和稳定表达GFP/MHBst155融合蛋白的Hep G2/GFP-MHBst155作为实验细胞,细胞用5-氮-2′-脱氧胞苷(5-Aza-Cd R,20μmol/L浓度)处理。采用四甲基偶氮噻唑蓝(MTT)法检测细胞吸光度值A490,分析细胞生长增殖情况;流式细胞术检测细胞周期变化。结果 72 h时Hep G2细胞组和Hep G2/GFP细胞组的A490分别为(0.67±0.12)、(0.70±0.06),与Hep G2/GFP-MHBst155细胞组(1.82±0.09)比较,差异均有统计学意义(t=-27.05、-36.71,P值均<0.05),而Hep G2细胞组与Hep G2/GFP细胞组比较差异无统计学意义(t=-1.21,P=0.24)。流式细胞术检测显示Hep G2/GFP-MHBst155细胞组的G0/G1期细胞比例为(26.23±2.70),明显低于Hep G2细胞组(45.20±1.25)和Hep G2/GFP细胞组(41.83±2.14),差异均有统计学意义(t=11.03、7.84,P值均<0.05);而经5-氮-2′-脱氧胞苷(5-Aza-Cd R)处理的Hep G2/GFP-MHBst155细胞组G0/G1期的比例为(43.03±1.45),与未处理Hep G2/GFP-MHBst155细胞组比较,差异有统计学意义(t=9.49,P<0.05)。结论 MHBst155可能通过缩短Hep G2细胞生长的G0/G1期而加快细胞周期进程,从而促进肝癌细胞的生长,其机制可能与MHBst155蛋白诱导生长调控基因的异常甲基化有关。更多还原

【Abstract】 Objective To explore the effect of C-terminally truncated middle hepatitis B virus surface proteins(MHBst155) on cell cycle and growth of human hepatocellular carcinoma cells. Methods Hep G2 / GFP-MHBst155 cell line stably expressing the fusion protein of a green fluorescent protein(GFP) and MHBst155 was used in the experiment. Cell growth was evaluated by MTT method. Cell cycle was determined by flow cytometry in these cells with or without treatment of 5-aza-2′-deoxycytidine(5-Aza-Cd R, 20 μmol / L). Results The A490 value in Hep G2 / GFP-MHBst155 was 1.82±0.09 at 72 hours of culture, and there was significant difference compared with Hep G2 cell(0.67 ±0.12, t =-27.05, P <0.05) and Hep G2 / GFP cell(0.70 ±0.06,t=-36.71, P<0.05). FCM analysis showed the percentage of G0/ G1 phase of Hep G2 / GFP-MHBst155(26.23±2.70)%, was significant lower compared with Hep G2(45.20±1.25, t=11.03, P<0.05) and Hep G2 / GFP(41.83±2.14, t=7.84, P<0.05). Furthermore,in Hep G2 / GFP-MHBst155 cells with 5-Aza-Cd R treatment, the percentage of G0/ G1phase(43.03 ±1.45, t =9.49, P <0.05) was significantly increased. Conclusion MHBst155 protein can shorten the G0/ G1 phase of cell cycle, and improve the proliferation ability of cells. The mechanism may be associated with MHBst155 inducing abnormal methylation of cell growth regulatory genes.更多还原