HCMV感染对胶质瘤细胞U87自噬的影响

Effects and Significance of HCMV Infection on Autophagy in U87 Glioma Cell

研究人巨细胞病毒(HCMV)感染对神经胶质瘤U87细胞自噬的影响。通过观察微管相关蛋白1轻链3(LC3)、自噬相关基因Beclin1及其蛋白表达的变化,从而探讨HCMV与神经胶质瘤发生、发展的关系及意义。用HCMV AD169(MOI=5)感染神经胶质瘤U87细胞,同时将未感染HCMV的U87细胞作为对照组。分别在6、12、24、48 h用RT-PCR检测Beclin1的表达,Western-blot和免疫荧光检测Beclin1和LC3编码蛋白的表达,最后用CCK-8检测细胞的增殖活性。结果显示,HCMV感染的U87细胞LC3-II蛋白表达水平逐渐下降(P<0.05);同时,HCMV感染的U87细胞Beclin1基因及蛋白的表达水平也逐渐下降(P<0.01),且HCMV感染U87细胞增殖显著(P<0.01)。以上结果表明,HCMV感染抑制胶质瘤U87细胞自噬,并会引起Beclin1表达水平下调,进而导致胶质瘤细胞增殖。

The effects of human cytomegalovirus（ HCMV） infection in U87 glioma cell autophagy were studied. The expression changes of microtubule associated protein 1 light chain 3（ LC3）,autophagy related gene Beclinl and its protein were observed,and accordingly investigate the relation of genesis,development,and significance of glioma with HCMV. Glioma U87 cell was infected with HCMV strain AD169（ MOI = 5）,and set U87 cell without HCMV infection as a control group. RT-PCR was used to detect the changes of expression of Beclin1 after HCMV infection at 6h,12 h,24 h,and 48 h respectively; Western-blot and immunofluorescence were used to test the expression of Beclin1 and LC3 coded proteins. Finally,CCK-8 assay was used to observe the proliferation activity of U87 glioma. The results showed that,the expression level of LC3-II protein in HCMV infected U87 cell was gradually decreased（ P〈 0. 05）,meanwhile,the expression level of Beclin1 gene and its protein was also gradually decreased（ P〈 0. 01）,moreover,the HCMV infected U87 cell proliferated significantly（ P〈 0. 01）. The results indicated that HCMV infected and inhibited U87 glioma cell autophagy,and cause the expression level of Beclin1 to lower,and resulted in the glioma cell to proliferate.