|
|
Effect of LncRNA HCG18 on PD-L1 Expression in Nasopharyngeal Carcinoma Cells and CD8+ T Cell Cytotoxic Activity |
LIU Chunjiang, et al |
The Fifth People's Hospital of Xinjiang Uygur Autonomous Region, Xinjiang Urumqi 830000, China |
|
|
Abstract Objective: To investigate the regulatory mechanism of LncRNA HCG18 on the function of CD8+ T cells against nasopharyngeal carcinoma (NPC) cells. Methods: The expression of HCG18 in NPC tissues and paracancerous tissues was detected by qRT-PCR. The expression of HCG18 in human nasal mucosal epithelial cells (HNEpC) and NPC cell lines (CNE1, CNE2, HONE-1, HNE-2) was also detected. CNE1 cells were divided into si-HCG18 group, si-NC group, and si-HCG18+PD-L1 group; HNE-2 cells were divided into HCG18 group, Vector group, and HCG18+sh-PD-L1 group. CNE1 and HNE-2 cells in each group were co-cultured with CD8+ T cells for 24 h (si-HCG18+CD8+T group, si-NC+CD8+T group, si-HCG18+PD-L1+CD8+T group, HCG18+CD8+T group, Vector+CD8+T group, and HCG18+sh-PD-L1+CD8+T group). The concentrations of INF-γ, TNF-α, and IL-2 in the co-culture system were detected by enzyme-linked immunosorbent assay (ELISA), and the tumor cell lysis rate was detected by a cytotoxicity assay kit. Western blot was used to detect PD-L1 protein expression. Dual luciferase reporter gene assay was used to verify the targeting relationship between HCG18 and miR-20b-5p, and the targeting relationship between miR-20b-5p and PD-L1. Results: The expression of HCG18 in NPC tissues was significantly higher than that in paracancerous tissues, and the expression of HCG18 in CNE1, CNE2, HONE-1, and HNE-2 cells was significantly higher than that in HNEpC cells (P<0.05). The expression of PD-L1 in CNE1 cells of the si-HCG18 group was significantly lower than that in the si-NC group (P<0.05), and the expression of PD-L1 in HNE-2 cells of the HCG18 group was significantly higher than that in the Vector group (P<0.05). Compared with the si-NC+CD8+T group, the concentrations of INF-γ, TNF-α, and IL-2 in the co-culture system of the si-HCG18+CD8+T group were significantly increased (P<0.05), and the CNE1 cell lysis rate was significantly increased (P<0.05). Compared with the si-HCG18+CD8+T group, the concentrations of INF-γ, TNF-α, and IL-2 in the co-culture system of the si-HCG18+PD-L1+CD8+T group were significantly decreased (P<0.05), and the CNE1 cell lysis rate was significantly decreased (P<0.05). Compared with the Vector+CD8+T group, the concentrations of INF-γ, TNF-α, and IL-2 in the co-culture system of the HCG18+CD8+T group were significantly decreased (P<0.05), and the HNE-2 cell lysis rate was significantly decreased (P<0.05). Compared with the HCG18+CD8+T group, the concentrations of INF-γ, TNF-α, and IL-2 in the co-culture system of the HCG18+sh-PD-L1+CD8+T group were significantly increased (P<0.05), and the HNE-2 cell lysis rate was significantly increased (P<0.05). miR-20b-5p is a target gene of HCG18, and PD-L1 is a target gene of miR-20b-5p. Conclusion: HCG18 upregulates PD-L1 expression in NPC cells and suppresses the anti-tumor activity of CD8+ T cells.
|
|
|
|
|
[1] Liu J,Zeng Z,Wang D,et al.Minimally invasive surgery for early-stage nasopharyngeal carcinoma[J].Craniofac Surg,2022,33(8):834-837. [2] Sathasivam HP,Chew SYL,Kim WR,et al.Nasopharyngeal carcinoma in adolescent patients:a case series[J].Clin Otolaryngol,2022,47(3):486-490. [3] Khorkova O,Stahl J,Joji A,et al.Long non-coding RNA-targeting therapeutics:discovery and development update[J].Expert Opin Drug Discov,2023,18(9):1011-1029. [4] Xu JY,Wei XL,Wang YQ,et al.Current status and advances of immunotherapy in nasopharyngeal carcinoma[J].Ther Adv Med Oncol,2022,7(14):175-184. [5] Zou C,Wu S,Wei H,et al.LINC01355 contributes to malignant phenotype of oral squamous cell carcinoma and cytotoxic T cell infiltration via activating notch signaling pathway[J].Immunol Res,2021,7(27):1830790-1830801. [6] Du Z,Wang B,Tan F,et al.The regulatory role of LncRNA HCG18 in various cancers[J].Mol Med (Berl),2023,101(4):351-360. [7] Lin M,Zhang XL,You R,et al.Evolutionary route of nasopharyngeal carcinoma metastasis and its clinical significance[J].Nat Commun,2023,14(1):610-630. [8] Lv J,Wei Y,Yin JH,et al.The tumor immune microenvironment of nasopharyngeal carcinoma after gemcitabine plus cisplatin treatment[J].Nat Med,2023,29(6):1424-1436. [9] Zhou Z,Chen Y.LncRNA SNHG1 promotes nasopharyngeal carcinoma development via targeting miR-424-5p[J].Histol Histopathol,2023,38(8):953-963. [10] Balakittnen J,Weeramange CE,Wallace DF,et al.Noncoding RNAs in oral cancer[J].Wiley Interdiscip Rev RNA,2023,14(3):e1754-e1765. [11] Zhang C,Lv H,Zhang F,et al.LncRNA HCG18 facilitates melanoma progression by modulating miR-324-5p/CDK16 axis[J].Am Transl Res,2022,14(2):1246-1257. [12] Ai Y,Wu S,Gao H,et al.Repression of CRNDE enhances the anti-tumour activity of CD8+T cells against oral squamous cell carcinoma through regulating miR-545-5p and TIM-3[J].Cell Mol Med,2021,25(23):10857-10868. [13] Zhao J,Bang S,Furutani K,et al.PD-L1/PD-1 checkpoint pathway regulates hippocampal neuronal excitability and learning and memory behavior[J].Neuron,2023,111(17):2709-2726. [14] Taheri M,Safarzadeh A,Hussen BM,et al.LncRNA/miRNA/mRNA network introduces novel biomarkers in prostate cancer[J].Cells,2022,11(23):3776-3803. [15] Jiang K,Zou H.microRNA-20b-5p overexpression combing pembrolizumab potentiates cancer cells to radiation therapy via repressing programmed death-ligand 1[J].Bioengineered,2022,13(1):917-929. |
|
|
|