HPV16 E2蛋白对巨噬细胞的影响  

Effect of HPV16 E2 protein on macrophages

作　　者：陈杰[1] 李沐宜 CHEN Jie;LI Mu-Yi(Yueyang Vourionad Technical College,Yueyang,Hu'nan 414000,China)

机构地区：[1]岳阳职业技术学院,湖南岳阳414000 [2]岳阳市第一人民医院,湖南岳阳414000

出　　处：《中国妇幼保健》2020年第19期3682-3684,共3页Maternal and Child Health Care of China

基　　金：湖南省教育厅高校科研项目(14C1152)。

摘　　要：目的研究人乳头状瘤病毒16 (HPV16) E_2蛋白对巨噬细胞的影响。方法构建真核表达pEGFP-C1/DBD、pEGFP-C1/HPV16 E_2及pEGFP-C1/TAD载体,观察HeLa细胞、巨噬细胞内HPV16 E_2与DBD、TAD表达、定位,检测HeLa细胞培养上清液内肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平,应用流式细胞仪检测巨噬细胞凋亡率。结果GFP组、GFP-E_2组的细胞核、细胞质中均存在绿色荧光,GFP-DBD组仅细胞核中存在绿色荧光,GFP-TAD组仅细胞质中存在绿色荧光。巨噬细胞内融合蛋白的绿色荧光分布位置和HeLa细胞相同。GFP组、GFP-DBD组细胞培养上清液内TNF-α、IL-1β水平与空白对照组比较差异均无统计学意义(均P>0.05);GFP-E_2组、GFP-TAD组细胞培养上清液内TNF-β、IL-1β水平显著高于空白对照组和GFP组(均P<0.05);GFP-TAD组细胞培养上清液内TNF-α水平高于GFP-E_2组(P<0.05)。GFP组、GFP-DBD组巨噬细胞凋亡率均略低于空白对照组,但差异均无统计学意义(均P>0.05);GFP-E_2、GFP-TAD组巨噬细胞凋亡率高于空白对照组和GFP组(均P<0.05);GFP-TAD组巨噬细胞凋亡率高于GFP-E_2组,差异有统计学意义(P<0.05)。结论 HeLa细胞、巨噬细胞内均能检出HPV16 E_2与TAD、DBD、GFP融合蛋白的真核表达载体,其中GFP-TAD主要分布在细胞质内,GFP-DBD主要分布在细胞核内;同时,HPV16 E_2与TAD高表达能使巨噬细胞释放TNF-β、IL-1上调,进而对巨噬细胞凋亡进行有效诱导。Objective To study the effect of human papillomavirus 16(HPV16)E2 on macrophages.Methods The eukaryotic expression vectors of pEGFP-C1/DBD,pEGFP-C1/HPV16 E2,pEGFP-C1/TAD were constructed.The expressions and locations of HPV16 E2,DBD,TAD in HeLa cells and macrophages were observed.The levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1(3)in culture supernatant of HeLa cells were detected.The rate of macrophages was determined by flow cytometry.Results Green fluorescence was observed in cell nucleus and cytoplasm in GFP group and GFP-E2 group;green fluorescence was observed in cell nucleus in GFP-DBD group;green fluorescence was observed in cytoplasm in GFP-TAD group.The distribution and location of green fluorescence of fusion protein in macrophages were similar to those in HeLa cells.There was no statistically significant difference in the levels of TNF-αand IL-1βin culture supernatant between GFP group,GFP-DBD group and blank control group(P>0.05))the levels of TNF-αand IL-1βin culture supernatant in GFP-E2 group and GFP-TAD group were statistically significantly higher than those in blank control group and GFP group(P<0.05))the level of TNF-αin culture supernatant in GFP-TAD group was statistically significantly higher than that in GFP-E2 group(P<0.05).The apoptotic rates of macrophages in GFP group and GFP-DBD group were lower than that in blank control group,but there was no statistically significant difference(P>0.05);the apoptotic rates of macrophages in GFP-E2 group and GFP-TAD group were statistically significantly higher than those in blank control group and GFP group(P<0.05);the apoptotic rate of macrophages in GFP-TAD group was statistically significantly higher than that in GFP-E2 group(P<0.05).Conclusion The eukaryotic expression vectors of HPV16 E2,TAD,DBD,GFP fusion proteins can be detected in HeLa cells and macrophages.GFP-TAD mainly locates in cytoplasm,and GFPDBD mainly locates in cell nucleus;the high expressions of HPV16 E2 and TAD can upregulate TNF-αand IL-1 and effectiv

关 键 词：人乳头状瘤病毒16 E_2蛋白 巨噬细胞 

分 类 号：R737.33[医药卫生—肿瘤]