人疱疹病毒6B感染对Molt3细胞增殖和细胞周期的影响  

Effects of human herpesvirus 6B infection on cell proliferation and cell cycle of Molt3 cells

作　　者：周锋[1] 冯东举[1] 顾斌[2] 姚堃[1] 李猛[1] 王金凤[1] 李凌云[3] 

机构地区：[1]南京医科大学微生物与免疫学系,江苏南京210029 [2]东南大学中大医院神经外科,江苏南京210009 [3]南京医科大学发育遗传学系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2014年第11期1457-1462,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助(81273235;81201520;81301698)

摘　　要：目的 :研究人疱疹病毒(human herpesvirus,HHV)6B感染T淋巴细胞系Molt3后对其细胞增殖和周期的影响。方法 :HHV-6B感染Molt3细胞后,倒置显微镜观察细胞形态变化,PCR鉴定Molt3细胞中HHV-6 U22基因,间接免疫荧光及Western blot检测HHV-6蛋白表达;MTT检测细胞增殖,流式细胞仪检测细胞周期变化,荧光定量PCR检测细胞周期相关蛋白m RNA水平变化。结果:HHV-6B感染48 h后,Molt3细胞出现典型细胞病变。PCR检测到感染组细胞中含有HHV-6 U22基因,间接免疫荧光及Western blot均检测到HHV-6蛋白表达。MTT显示HHV-6B能明显抑制Molt3细胞的增殖。HHV-6B感染后Molt3细胞周期发生改变,与对照组相比,感染组细胞G1期增多,S期和G2期减少。实时荧光定量PCR表明HHV-6B感染后cyclin E1m RNA水平降低,p53 m RNA水平升高。结论:HHV-6B能够有效感染Molt3细胞引起典型的细胞病变效应,HHV-6B感染使细胞周期阻滞在G1期从而抑制细胞增殖。Objective：To study the significance of human herpesvirus type 6B（HHV-6B） infection on human T-lymphoblastoid cell line Molt3 and its effects on cell cycle and proliferation. Methods：HHV-6B-infected Molt3 cell morphology was assessed by a inverted microscope. The fragment of HHV-6 U22 gene in Molt3 cells was amplified by PCR. Expression of HHV-6B late protein was examined by the immunofluorescence assay and Western Blot. Cell proliferation was measured by MTT assay after HHV-6B infected Molt3. Cell cycle analysis was evaluated by flow cytometry. The m RNA levels of cell cycle related protein were examined by Realtime PCR. Results：Molt3 cells infected by HHV-6B showed typical cytopathic effect at 48 h post-infection. HHV-6 U22 gene was detected by PCR. HHV-6 late protein was positive in infected cells by indirect immunofluorescence and Western Blot assay. MTT assay detected that HHV-6B infection inhibited the proliferation of Molt3 cells significantly. Compared with the uninfected cells,percentages of Molt3 cells infected by HHV-6B were increased in phase G1 but decreased in phase S and G2. The m RNA levels of cyclin E1 were decreased after 48 h post-infection,however,the m RNA levels of p53 were significantly increased after infection.Conclusion：HHV-6B can infect the Molt3 cells,lead to typical cytopathic effect,and inhibit cell proliferation and make the cell cycle stay in phase G1.

关 键 词：人疱疹病毒6B 细胞增殖 细胞周期 

分 类 号：R371.11[医药卫生—病原生物学]