circNRIP1 induces CD8+T cells by upregulating PD-L1 expression in cervical cancer cells
10.3969/j.issn.1673-4130.2025.12.014
- VernacularTitle:circNRIP1上调宫颈癌细胞PD-L1表达诱导CD8+T细胞的耗竭研究
- Author:
Lingling YAN
1
;
Xiaoxia ZHANG
;
Xiaoli CAO
Author Information
1. 南通市肿瘤医院检验科,江苏南通 226000
- Keywords:
cervical cancer;
circNRIP1;
CD8+T cells;
exhaustion
- From:
International Journal of Laboratory Medicine
2025;46(12):1485-1491
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect and mechanism of circular RNA nuclear receptor interacting protein 1(circNRIP1)expression on exhaustion of CD8+T cells in cervical cancer cells.Methods Real-time quantitative PCR was used to detect the expression of circNRIP1 in cervical epithelial cells HCerEpic and cer-vical cancer cells C-33A,Hela,SiHa,and CS121.C-33A cells were divided into Vector group,circNRIP1 group,and circNRIP1+miR-138-5p group,while CS121 cells were divided into sh-NC group,sh-circNRIP1 group,and sh-circNRIP1+miR-138-5p inhibitor.Human peripheral blood CD8+T cells were extracted,and C-33A cells in Vector group and circNRIP1 group were co-incubated with CD8+T cells for 24 hours(CD8+T/Vector group and CD8+T/Vector group).CS121 cells in sh-NC group and sh-circNRIP1 group were co-incu-bated with CD8+T cells for 24 hours(CD8+T/sh NC group and CD8+T/sh-circNRIP1 group).Cytotoxicity experiments were conducted to detect the killing ability of CD8+T cells,ELISA was used to detect the levels of interleukin(IL)-2,interferon(IFN)-γ,and tumor necrosis factor(TNF)-α in the cell supernatant.Flow cy-tometry was used to detect the expression of programmed death receptor-1(PD-1),T cell Immunoglobulin domain and Mucin domain protein-3(TIM3),and lymphocyte activation gene 3(LAG3)in CD8+T cells.Dual luciferase reporter gene experiments were conducted to verify the targeting relationship between circNRIP1 and miR-138-5p,as well as the targeting relationship between miR-138-5p and PD-L1.Results The expres-sions of circNRIP1 in C-33A,Hela,SiHa,and CS121 cells were significantly higher than those in HCerEpic(P＜0.05).The killing ability of CD8+T cells against C-33A cells in the circNRIP1 group was lower than their killing ability against Vector group cells.The levels of IL-2,IFN-γ,and TNF-α secreted by CD8+T cells in the CD8+T/circNRIP1 group were significantly lower than those in the CD8+T/Vector group(P＜0.05),and the levels of PD-1,LAG-3,and TIM-3 expressed by CD8+T cells in the CD8+T/circNRIP1 group were al-so significantly higher than those in the CD8+T/Vector group(P＜0.05).The killing ability of CD8+T cells against sh-circNRIP1 group CS121 cells was higher than their killing ability against sh-NC group cells(P＜0.05).The levels of IL-2,IFN-γ,and TNF-α secreted by CD8+T cells in the CD8+T/sh-circNRIP1 group were significantly higher than those in the CD8+T/sh-NC group(P＜0.05).The PD-1,LAG-3,and TIM-3 levels of CD8+T cells in the CD8+T/sh-circNRIP1 group were also significantly lower than those in the CD8+T/sh-NC group(P＜0.05).The results of the dual-luciferase reporter gene experiment showed that miR-138-5p was the target gene of circNRIP1,and PD-L1 was the target gene of miR-138-5p.Conclusion circNRIP1 can in-duce the exhaustion of CD8+T cells by upregulating the expression of PD-L1.
