下调USP37抑制PD-L1表达增强三阴性乳腺癌细胞免疫反应-现代医学

下调USP37抑制PD-L1表达增强三阴性乳腺癌细胞免疫反应

单位：1. 青岛大学附属青岛市市立医院/青岛市市立医院肿瘤科, 山东青岛 266071;
2. 中国人民解放军海军第九七一医院肿瘤科, 山东青岛 266071

分类号：R737.9

出版年·卷·期（页码）：2024·52·第十二期（1863-1869）

摘要：

目的：探讨下调USP37表达对三阴性乳腺癌(TNBC)细胞程序性死亡受体配体1(PD-L1)表达量的影响及对其抗肿瘤免疫反应的作用。方法：应用免疫组织化学染色法检测TNBC组织中USP37的表达量及其与PD-L1表达量之间的相关性。反用siRNA瞬时转染技术降低人TNBC细胞系MDA-MB-231细胞中USP37的表达。采用实时定量聚合酶链反应(qRT-PCR)、蛋白质印迹法(Western Blot)检测瞬时转染后MDA-MB-231细胞中USP37和PD-L1的mRNA及蛋白质的表达水平。将瞬时转染后的MDA-MB-231细胞与外周血单个核细胞(PBMC)进行共培养，采用CCK8法评估PBMC对MDA-MB-231细胞的杀伤作用，流式细胞术检测MDA-MB-231细胞中PD-L1的表达变化。结果：免疫组化结果显示，TNBC中USP37高表达且与PD-L1的表达量呈正相关(P＜0.01)。与转染对照组相比，下调USP37后MDA-MB-231细胞中USP37和PD-L1的mRNA、蛋白质表达量均显著降低。CCK8实验显示，下调USP37增强PBMC对MDA-MB-231细胞的杀伤能力(P＜0.01)；流式细胞术显示，下调USP37 MDA-MB-231细胞表面PD-L1表达量下调(P＜0.01)。结论：下调USP37可抑制MDA-MB-231细胞PD-L1的表达，影响TNBC的抗肿瘤免疫反应。

Objective:To investigate the effect of downregulating USP37 expression on programmed death-ligand 1(PD-L1)expression in triple-negative breast cancer(TNBC) cells and its role in tumor immune response. Methods: Immunohistochemistry was used to detect the expression of USP37 and its correlation with PD-L1 expression in TNBC tissues. siRNA-mediated transient transfection was performed to downregulate USP37 expression in MDA-MB-231 cells. qRT-PCR and Western Blot were used to measure the mRNA and protein levels of USP37 and PD-L1 in MDA-MB-231 cells after transient transfection. MDA-MB-231 cells transfected with siRNA were co-cultured with peripheral blood mononuclear cells(PBMCs), and CCK8 assay was used to evaluate the cytotoxic effect of PBMCs on MDA-MB-231 cells. Flow cytometry was used to detect changes in PD-L1 expression on MDA-MB-231 cells. Results: Immunohistochemistry showed that USP37 was highly expressed in TNBC tissues and positively correlated with PD-L1 expression(P<0.01). Compared to the control group, downregulation of USP37 significantly reduced the mRNA and protein levels of both USP37 and PD-L1 in MDA-MB-231 cells. CCK8 assay indicated that downregulation of USP37 enhanced PBMC-mediated cytotoxicity against MDA-MB-231 cells(P<0.01). Flow cytometry showed that the expression of PD-L1 on the surface of MDA-MB-231 cells was significantly reduced in the USP37 downregulation group(P<0.01).Conclusion: Downregulation of USP37 inhibits PD-L1 expression on MDA-MB-231 cells and influences the anti-tumor immune response in TNBC.

参考文献：

[1] SUNG H,FERLAY J,SIEGEL R L,et al.Global cancer statistics 2020:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J].CA Cancer J Clin,2021,71(3):209-249.
[2] ONKAR S S,CARLETON N M,LUCAS P C,et al.The great immune escape:understanding the divergent immune response in breast cancer subtypes[J].Cancer Discov,2023,13(1):23-40.
[3] WAKS A G,WINER E P.Breast cancer treatment:a review[J].JAMA,2019,321(3):288-300.
[4] SCHMID P,CORTES J,PUSZTAI L,et al.Pembrolizumab for early triple-negative breast cancer[J].N Engl J Med,2020,382(9):810-821.
[5] QIN S,XU L,YI M,et al.Novel immune checkpoint targets:moving beyond PD-1 and CTLA-4[J].Mol Cancer,2019,18(1):155.
[6] PUSZTAI L,KARN T,SAFONOV A,et al.New strategies in breast cancer:immunotherapy[J].Clin Cancer Res,2016,22(9):2105-2110.
[7] QIN G,WANG X,YE S,et al.NPM1 upregulates the transcription of PD-L1 and suppresses T cell activity in triple-negative breast cancer[J].Nat Commun,2020,11(1):1669.
[8] CHAUHAN R,BHAT A A,MASOODI T,et al.Ubiquitin-specific peptidase 37:an important cog in the oncogenic machinery of cancerous cells[J].J Exp Clin Cancer Res,2021,40(1):356.
[9] MANCZYK N,VEGGIANI G,TEYRA J,et al.The ubiquitin interacting motifs of USP37 act on the proximal Ub of a di-Ub chain to enhance catalytic efficiency[J].Sci Rep,2019,9(1):4119.
[10] WU C,CHANG Y,CHEN J,et al.USP37 regulates DNA damage response through stabilizing and deubiquitinating BLM[J].Nucleic Acids Res,2021,49(19):11224-11240.
[11] QIN T,LI B,FENG X,et al.Correction to:abnormally elevated USP37 expression in breast cancer stem cells regulates stemness,epithelial-mesenchymal transition and cisplatin sensitivity[J].J Exp Clin Cancer Res,2021,40(1):354.
[12] GARRIDO-CASTRO A C,LIN N U,POLYAK K.Insights into molecular classifications of triple-negative breast cancer:improving patient selection for treatment[J].Cancer Discov,2019,9(2):176-198.
[13] KUDELOVA E,SMOLAR M,HOLUBEKOVA V,et al.Genetic heterogeneity,tumor microenvironment and immunotherapy in triple-negative breast cancer[J].Int J Mol Sci,2022,23(23):14937.
[14] CHEN C,LI S,XUE J,et al.PD-L1 tumor-intrinsic signaling and its therapeutic implication in triple-negative breast cancer[J].JCI Insight,2021,6(8):e131458.
[15] CASEY S C,TONG L,LI Y,et al.MYC regulates the antitumor immune response through CD47 and PD-L1[J].Science,2016,352(6282):227-231.
[16] XIAO X,SHI J,HE C,et al.ERK and USP5 govern PD-1 homeostasis via deubiquitination to modulate tumor immunotherapy[J].Nat Commun,2023,14(1):2859.
[17] XIONG W,GAO X,ZHANG T,et al.USP8 inhibition reshapes an inflamed tumor microenvironment that potentiates the immunotherapy[J].Nat Commun,2022,13(1):1700.
[18] YANG Z,XU G,WANG B,et al.USP12 downregulation orchestrates a protumourigenic microenvironment and enhances lung tumour resistance to PD-1 blockade[J].Nat Commun,2021,12(1):4852.
[19] ZOU Q,JIN J,XIAO Y,et al.T cell intrinsic USP15 deficiency promotes excessive IFN-γ production and an immunosuppressive tumor microenvironment in MCA-induced fibrosarcoma[J].Cell Rep,2015,13(11):2470-2479.
[20] YANG S,YAN H,WU Y,et al.Deubiquitination and stabilization of PD-L1 by USP21[J].Am J Transl Res,2021,13(11):12763-12774.
[21] LI Q,ZHANG L,YOU W,et al.PRDM1/BLIMP1 induces cancer immune evasion by modulating the USP22-SPI1-PD-L1 axis in hepatocellular carcinoma cells[J].Nat Commun,2022,13(1):7677.
[22] WU J,GUO W,WEN D,et al.Deubiquitination and stabilization of programmed cell death ligand 1 by ubiquitin-specific peptidase 9,X-linked in oral squamous cell carcinoma[J].Cancer Med,2018,7(8):4004-4011.
[23] QIN T,CUI X Y,XIU H,et al.USP37 downregulation elevates the chemical sensitivity of human breast cancer cells to adriamycin[J].Int J Med Sci,2021,18(2):325-334.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录