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目的:研究人类疱疹病毒6型(HHV-6)感染人星形胶质瘤细胞U251,及感染后对其生物学功能的影响。方法:HHV-6感染U251细胞建立HHV-6体外感染模型,倒置显微镜观察细胞病变。PCR法检测HHV-6u22基因。间接免疫荧光(IFA)检测HHV-6即刻早期蛋白(IE1)和晚期蛋白(gB)的表达。MTT法检测U251感染HHV-6后细胞增殖的改变。流式细胞术(FCM)检测U251细胞周期改变。结果:在感染3天后U251出现典型细胞病变,细胞肿胀增大数倍,透亮,呈多形性。PCR检测到HHV-6u22基因。IFA检测到HHV-6IE1蛋白和gB蛋白的表达。MTT法显示HHV-6能增强U251细胞的增殖能力。FCM检测表明,HHV-6感染后U251细胞周期发生改变,感染组和对照组相比,前者G1期的细胞减少,S期和G2期细胞增多。结论:HHV-6能感染U251细胞引起细胞病变,促进细胞的增殖,使细胞周期发生改变。本研究结果提示人类疱疹病毒6型可能参与人神经胶质瘤的发生、发展,对其机制尚不清楚。
Objective: To study the effect of human herpesvirus type 6 (HHV-6) on human astrocytic glioma U251 and its biological function after infection. Methods: Infection of U251 cells with HHV-6 was used to establish a model of HHV-6 infection in vitro. The cytopathic effect was observed by inverted microscope. HHV-6u22 gene was detected by PCR. Indirect immunofluorescence (IFA) was used to detect the expression of HHV-6 immediate early protein (IE1) and late glycoprotein (gB). MTT assay U251 infected with HHV-6 cell proliferation changes. Cell cycle changes of U251 cells were detected by flow cytometry (FCM). Results: U251 cells showed typical cytopathic effect after 3 days of infection, the cell swelling increased several times, translucent and pleomorphic. HHV-6u22 gene was detected by PCR. IFA detected the expression of HHV-6IE1 protein and gB protein. MTT assay showed that HHV-6 enhanced the proliferation of U251 cells. The results of FCM showed that the cell cycle of U251 cells was changed after HHV-6 infection. Compared with the control group, the number of cells in G1 phase and the cells in S phase and G2 phase increased in infected group. Conclusion: HHV-6 can infect U251 cells to cause cytopathic effect, promote cell proliferation and change cell cycle. The results suggest that human herpes virus type 6 may be involved in the occurrence and development of human glioma, the mechanism is unclear.