circRNA-1565抑制CD8^(+)T细胞对宫颈癌细胞的杀伤功能研究  

circRNA-1565 inhibits function of CD8^(+)T cells and regulates its killing function to cervical cancer cells

作　　者：王青慧 李波 王晓黎 胡传翠 聂明朝 庞晓庆 张亚芳 WANG Qinghui;LI Bo;WANG Xiaoli;HU Chuancui;NIE Mingchao;PANG Xiaoqing;ZHANG Yafang(Department of Obstetrics and Gynecology,Hainan Women and Children's Medical Center,Haikou 570206,China)

机构地区：[1]海南省妇女儿童医学中心妇产科,海口570206

出　　处：《中国免疫学杂志》2023年第10期2191-2195,共5页Chinese Journal of Immunology

基　　金：海南省科学技术厅项目(821RC715);海南省临床医学中心建设项目(琼卫医函【2021】75号)资助。

摘　　要：目的:探究circRNA-1565对CD8^(+)T细胞的作用及其对宫颈癌细胞杀伤能力的影响。方法:磁珠分选并用流式细胞术鉴定CD8^(+)T细胞。所得CD8^(+)T细胞分为Control组(转染空白质粒)和circRNA-1565组(转染circRNA-1565质粒)。CCK-8和流式细胞术检测Control组和circRNA-1565组CD8^(+)T细胞增殖能力和凋亡水平,Western blot检测Control组和circRNA-1565组CD8+T细胞Bcl-2、Bax和cleaved-caspase-3表达。将HeLa和SiHa细胞分别与Control组和circRNA-1565组CD8^(+)T细胞共孵育(HeLa+CD8^(+)T+Vector组、HeLa+CD8+T+circRNA-1565组、SiHa+CD8^(+)T+Vector组、SiHa+CD8+T+circRNA-1565组),检测各组CD8^(+)T细胞对HeLa和SiHa细胞的杀伤能力,ELISA检测共孵育后各组CD8^(+)T细胞分泌TNF-α、IFN-γ、Granzyme-B和perforin的能力。结果:磁珠分选的人PBMC CD8^(+)T细胞阳性率>88%。相比于Control组,circRNA-1565组CD8^(+)T细胞增殖能力显著下降(P<0.001),细胞凋亡显著增加(P<0.001),Bcl-2和Bax表达显著降低(P<0.001),cleaved-caspase-3表达显著增加。相比于HeLa+CD8+T+Vector组和SiHa+CD8^(+)T+Vector组,HeLa+CD8^(+)T+circRNA-1565组以及SiHa+CD8^(+)T+circRNA-1565组细胞上清中TNF-α、IFN-γ、Granzyme-B和perforin浓度均显著降低(P<0.001),对Hela和SiHa细胞杀伤能力显著降低(P<0.001)。结论:circRNA-1565调控CD8^(+)T细胞增殖和凋亡从而抑制其对宫颈癌细胞的杀伤能力。Objective:To explore role of circRNA-1565 in CD8^(+)T cells and its effect on killing ability of cervical cancer cells.Methods:Magnetic bead was used to sort and flow cytometry was used to identify CD8+T cells.CD8+T cells obtained were divided into Control group(transfected with blank plasmid)and circRNA-1565 group(transfected with circRNA-1565 plasmid).Proliferation ability and apoptosis level of CD8^(+)T cells in Control group and circRNA-1565 group were detected by CCK-8 and flow cytometry.Western blot was used to detect expressions of Bcl-2,Bax and cleaved-caspase-3 of CD8^(+)T cells in Control group and circRNA-1565 group.HeLa and SiHa cells were co-incubated with CD8^(+)T cells in Control group and circRNA-1565 group(HeLa+CD8^(+)T+Vector group,HeLa+CD8^(+)T+circRNA-1565 group,SiHa+CD8^(+)T+Vector group,SiHa+CD8^(+)T+circRNA-1565 group),and killing ability of CD8^(+)T cells on HeLa and SiHa cells in each group were detected.After co-incubation,ELISA was used to detect ability of CD8^(+)T cells to secrete TNF-α,IFN-γ,Granzyme-B and perforin.Results:Positive rate of CD8^(+)T cells in human PBMCs sorted by magnetic beads was more than 88%.Compared with control group,proliferation ability of CD8^(+)T cells in circRNA-1565 group was significantly decreased(P<0.001),apoptosis was significantly increased(P<0.001),expressions of Bcl-2 and Bax were significantly decreased(P<0.001),and expression of cleaved-caspase-3 was significantly increased(P<0.001).Compared with HeLa+CD8^(+)T+Vector group or SiHa+CD8^(+)T+Vector group,HeLa+CD8^(+)T+circRNA-1565 group and SiHa+CD8^(+)T+circRNA-1565 group had significantly lower concentrations of TNF-α,IFN-γ,Granzyme-B and perforin inCD8^(+)T cells(P<0.001),killing abilities to HeLa cells and SiHa cells were significantly decreased(P<0.001).Conclusion:circRNA-1565 regulates proliferation and apoptosis of CD8+T cells and inhibits their ability to kill cervical cancer cells.

关 键 词：circRNA-1565 CD8^(+)T 宫颈癌 杀伤功能 

分 类 号：R392.12[医药卫生—免疫学]