人巨细胞病毒感染对HEL细胞周期及Cdt1表达的影响

Influence of human cytomegalovirus infection on cell cycle and replication licensing factor Cdt1 in human embryonic lung fibroblastic cells

摘要:

目的：研究人巨细胞病毒（HCMV）感染对人胚肺成纤维（HEL）细胞周期及复制允许因子Cdt1表达的影响，探讨HCMV感染可能的发病机制。方法：用血清饥饿法同步化HEL细胞于G0/G1期, 用HCMV感染同步化的HEL细胞作为感染组（n=106），同时设模拟感染组为对照组（n=106）。于感染后12，24，48，72，96 h收获细胞。流式细胞术测细胞周期； RT-PCR法检测Cdt1 mRNA的表达水平。结果：①模拟感染组在感染后12 h，24 h 时处于G1期细胞比感染组明显增多，两组比较，差异有显著性（P﹤0.01）。②两组Cdt1 mRNA表达水平在12 h，24 h和48 h时差异均有显著性（P﹤0.05）。模拟感染组Cdt1 mRNA表达水平的高峰出现在感染后12 h，而感染组的高峰则出现在感染后48 h。结论： ①HCMV感染影响了HEL细胞周期，使大部分细胞停滞在G1期。②HCMV感染使HEL细胞复制允许因子Cdt1 mRNA的表达水平迟滞。③HCMV感染可能通过影响复制允许因子Cdt1的生成而干扰宿主细胞周期，影响宿主细胞DNA合成，导致器官功能障碍。［中国当代儿科杂志，2007，9（6）：580-582］

Abstract:

OBJECTIVE: To study the influence of human cytomegalovirus (HCMV) infection on cell cycle and the expression of replication licensing factor Cdt1 in human embryonic lung fibroblastic (HEL) cells and to explore the pathogenesis of HCMV infection. MethodsHEL cells were synchronized in the G0/G1 phase by the serum starvation method. The synchronized HEL cells were infected with HCMV, and those that were not subjected to HCMV infection were used as the control group. The HEL cells were harvested at 12, 24, 48, 72 and 96 hrs of HCMV infection. The cell cycle of HEL cells was detected by the flow cytometry. The expression of Cdt1 mRNA in HEL cells was determined by reverse transcription-ploymerase chain reaction (RT-PCR). ResultsThe cells in the G1 phase in the control group was significantly more than in the HCMV-infected group 12 and 24 hrs after infection (P<0.01). The expression of Cdt1 mRNA in the HCMV-infected group was significantly lower 12 and 24 hrs after infection but increased significantly 48 hrs after infection compared with the control group (P<0.05). The expression of Cdt1 mRNA reached a peak at 12 hrs of infection in the control group, but at 48 hrs of infection in the HCMV-infected group, which markedly lagged behind the control group.ConclusionsHCMV infection arrests the cell cycle of HEL cells at the G1 phase. HCMV infection makes Cdt1 expression delay. HCMV infection can interfere cell cycle of HEL cells possibly through affecting the expression of Cdt1.［Chin J Contemp Pediatr, 2007, 9 (6):580-582］