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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。
Objective To investigate the effect of MHC-155 on the growth and proliferation of hepatocellular carcinoma cells. Methods Hep G2 cells and Hep G2 / GFP-MHBst155 stably expressing GFP / MHBst155 fusion protein were used as experimental cells and cells were treated with 5-Aza-Cd (20 μmol / L) . Cell absorbance value A490 was detected by MTT assay, cell growth and proliferation were analyzed, and cell cycle changes were detected by flow cytometry. Results The A490 of Hep G2 group and Hep G2 / GFP group at 72 h were (0.67 ± 0.12) and (0.70 ± 0.06), respectively, which were significantly lower than those of Hep G2 / GFP-MHBst155 group (1.82 ± 0.09) The statistical significance (t = -27.05, -36.71, P <0.05). There was no significant difference between Hep G2 cells and Hep G2 / GFP cells (t = -1.21, P = 0.24). Flow cytometry showed that the percentage of G0 / G1 phase cells in Hep G2 / GFP-MHBst155 cells was (26.23 ± 2.70), significantly lower than that in Hep G2 cells (45.20 ± 1.25) and Hep G2 / GFP cells (41.83 ± 2.14). The differences were statistically significant (t = 11.03, 7.84, P <0.05). However, Hep G2 / GFP- The proportion of G0 / G1 phase in MHBst155 cells was (43.03 ± 1.45), which was significantly different from that in untreated Hep G2 / GFP-MHBst155 cells (t = 9.49, P <0.05). Conclusions MHBst155 may accelerate the cell cycle progression by shortening the G0 / G1 phase of Hep G2 cell growth, which may be related to the abnormal methylation induced by MHBst155 protein.