CAJ | 학술논문

目的研究人巨细胞病毒(human cytomegalovirus,HCMV)感染对人胚肺成纤维(human embryonic lung fibroblasts,HEL)细胞Cdtl与Geminin因子表达的影响.方法用血清饥饿法同步化HEL细胞于G0/G1期,流式细胞术测细胞周期;用HCMV感染同步化的HEL细胞作为感染组,同时设模拟感染组为对照组.于感染后12、24、48、72和96 h收获细胞,提取总的RNA,采用半定量逆转录-聚合酶链反应检测Cdtl-mRNA和Geminin-mRNA的表达水平.结果 (1)血清饥饿后的HEL细胞82.8%处于G0/G1期.(2)12、24、48和72 h两组HEL细胞Cdtl mRNA表达水平比较均有统计学意义(t值分别为32.231,6.349,-5.846,-1.573;P均为0.000),12和24 h时感染组较模拟感染组明显降低,48和72 h时感染组较模拟感染组明显升高.(3)12、24、48和72 h两组HEL细胞Geminin mRNA表达水平比较均有统计学意义(t值分别为-8.891,-6.483,13.010,10.572;P均为0.000),12和24 h时感染组较模拟感染组明显增加,48和72 h时模拟感染组较感染组明显升高.结论 (1)血清饥饿法将绝大部分细胞同步于G0/G1期,血清饥饿法同步化HEL细胞效果较佳.(2)HCMV能诱导Cdtl及Geminin因子表达异常,使Geminin/Cdtl平衡失调,这可能是HCMV感染导致细胞周期阻滞于G1期的机制.
목적 연구인거세포병독(human cytomegalovirus,HCMV)감염대인배폐성섬유(human embryonic lung fibroblasts,HEL)세포Cdtl여Geminin인자표체적영향.방법 용혈청기아법동보화HEL세포우G0/G1기,류식세포술측세포주기;용HCMV감염동보화적HEL세포작위감염조,동시설모의감염조위대조조.우감염후12、24、48、72화96 h수획세포,제취총적RNA,채용반정량역전록-취합매련반응검측Cdtl-mRNA화Geminin-mRNA적표체수평.결과 (1)혈청기아후적HEL세포82.8%처우G0/G1기.(2)12、24、48화72 h량조HEL세포Cdtl mRNA표체수평비교균유통계학의의(t치분별위32.231,6.349,-5.846,-1.573;P균위0.000),12화24 h시감염조교모의감염조명현강저,48화72 h시감염조교모의감염조명현승고.(3)12、24、48화72 h량조HEL세포Geminin mRNA표체수평비교균유통계학의의(t치분별위-8.891,-6.483,13.010,10.572;P균위0.000),12화24 h시감염조교모의감염조명현증가,48화72 h시모의감염조교감염조명현승고.결론 (1)혈청기아법장절대부분세포동보우G0/G1기,혈청기아법동보화HEL세포효과교가.(2)HCMV능유도Cdtl급Geminin인자표체이상,사Geminin/Cdtl평형실조,저가능시HCMV감염도치세포주기조체우G1기적궤제.
Objective To determine the expressions of Cdtl and Oeminin genes in human embryonic lung fibroblasts(HEL) and the influence of human cytomegalovirus(HCMV)infection.Methods The HEL cells were synchronized in G0/G1 by serum starvation.Cell cycle of HEL cells was detected by flow cytometry.Synchronized HEL cells were infected with HCMV,at the same time the mock-infected group was set as control.Infected HEL cells were harvested at 12 h,24 h,48 h,72 h and 96 h post infection.The expressions of Cdtl and Geminin genes were detected with semi.quantitative RT-PCR. Results 82.8% of the HEL cells were in G0/G1 phase after serum starvation.Analysis showed that the expressions of Cdtl gene in HCMV-infected group were statistically lower(all P=0.000) than those in the mock-infected group at 12 h and 24 h post infection,but higher than the latter group at 48 h and 72 h post irrfection.The expressions of Geminin mRNA in HCMV-infected group were statistically higher than those of the mock-infected group at 12 h and 24 h post infection,but lower than tha latter group at 48 h and 72 h post infection(all P=0.000). Conclusions Serum starvation is a better method to synchronize the HEL cell.HCMV can induce abnormal expressions of Cdtl and Geminin genes and Geminin/Cdtl imbalance.which may play a role in HCMV blocked cell cycle in G1 phase.