HLA-G通过促进活化性受体的表达增强NK细胞对宫颈癌细胞的杀伤作用  

HLA-G Enhances the Killing Effect of NK Cells on Cervical Cancer Cells by Promoting the Expression of Activated Receptor

作　　者：刘倩 哈提拉·吐尔逊[1] 阿仙姑·哈斯木[1] LIU Qian;HATILA Tuerxun;AXIANGU Hasimu(Xinjiang Key Laboratory of Molecular Biology of Endemic Diseases,School of Basic Medicine,Xinjiang Medical University,Urumqi Xinjiang 830054,China)

机构地区：[1]新疆医科大学基础医学院,新疆地方病分子生物学重点实验室,新疆乌鲁木齐830054

出　　处：《昆明医科大学学报》2022年第7期46-54,共9页Journal of Kunming Medical University

基　　金：新疆维吾尔自治区自然科学基金青年项目(2019D01C190)。

摘　　要：目的探索宫颈癌细胞中HLA-G表达差异对NK细胞表面活化性受体的影响。方法实时定量PCR(real-time PCR)及免疫印迹法(Western-Blot WB)检测宫颈癌细胞及正常宫颈细胞中HLA-G的表达差异。利用慢病毒载体构建HLA-G稳定下调的宫颈癌细胞株,并与NK细胞共培养,CCK8法检测共培养后宫颈癌细胞增殖能力的变化,流式细胞术检测宫颈癌细胞凋亡水平。同时,流式细胞术检测共培养后NK细胞表面活化性受体NKG2D、NKP30的表达水平及颗粒素(Granzyme)、穿孔酶(Perforin)的表达差异。结果RT-PCR及WB法检测结果显示:与H8细胞相比,C33a、SiHa细胞中HLA-G的表达显著升高(P<0.05)。CCK8法检测结果显示:HLA-G下调后,C33a及SiHa细胞与NK细胞共培养,C33a及SiHa细胞的增殖能力显著减弱(P<0.05),同时,C33a及SiHa细胞的凋亡水平显著升高(P<0.05)。流式细胞术检测结果显示:C33a及SiHa细胞HLAG下调后,与其共培养的NK表面NKG2D表达显著升高(P<0.05),与C33a共培养的NK细胞表面NKP30及Granzyme、Perforin表达显著升高(P<0.05),与SiHa共培养的NK细胞表面NKP30及Granzyme、Perforin表达存在升高趋势,但差异均无统计学意义(P>0.05)。结论HLA-G在宫颈癌细胞中高表达,HLA-G表达升高可促进活化性受体的表达,增强NK细胞对宫颈癌细胞的杀伤作用。Objective To explore the effect of HLA-G expression on NK cell surface activation receptor in cervical cancer cells.Methods Real-time PCR(RT-PCR)and Western Blot(WB)methods were used to examine the expression of HLA-G in cervical cancer cells and normal cervical cells,respectively.The cervical cancer cell lines transfected with lentivirus to silence the expression of HLA-G were co-cultured with NK cells.The proliferation ability of co-cultured cervical cancer cells was measured by the method of CCK8,and the level of apoptosis in cervical cancer cells was measured by the flow cytometry method,as well as the expression of NKG2D and NKP30,Granzyme and Perforin.Results The results of RT-PCR and WB showed that compared with H8 cells,the expressions of HLA-G in C33a and SiHa cells were significantly increased(P<0.05).Results of CCK8 showed that the proliferation of C33a and SiHa cells was significantly decreased(P<0.05)when C33a and SiHa cells were co-cultured with NK cells after transfection.The levels of apoptosis in C33a and SiHa cells were significantly increased(P<0.05).The results of Flow cytometry showed that the expression of NKG2D on NK surface was significantly increased after C33a and SiHa were transfected(P<0.05).Of C33a,The expression of NKP30 on the surface of NK cells and Granzyme,Perforin were significantly increased(P<0.05).However,of C33a,the expression of NKP30 on the surface of NK cells and Granzyme,Perforin were increased,but the difference was not statistically significant(P>0.05).Conclusion The high expression of HLA-G is in cervical cancer cells,and it may enhance the killing effect of NK cells on cervical cancer cells by promoting the expression of activated receptor.

关 键 词：宫颈癌 HLA-G NK细胞 免疫逃逸 

分 类 号：R730.3[医药卫生—肿瘤]