CCR5介导小鼠肝癌微环境中骨髓来源抑制性细胞的迁移
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摘要
目的探索CCR5在小鼠肝癌微环境中MDSCs迁移的作用。方法建立原位肝癌小鼠模型,流式检测其骨髓来源抑制性细胞(myeloid-derived suppressor cells, MDSCs)水平。分选MDSCs,用CCR5抑制剂100 nmol/L预处理后观察小鼠肝癌细胞H22条件培养基(tumor conditional medium, TCM)及CCR5抑制剂对MDSCs迁移的影响。最后在小鼠肝癌原位模型尾静脉注射DiR标记的CCR5抑制剂处理MDSCs,观察体内肝癌微环境下CCR5抑制剂对MDSCs的迁移影响。结果在原位肝癌小鼠模型中,MDSCs在骨髓、脾、肝中的水平与正常小鼠相比显著性升高;分选得到MDSCs纯度可达94.5%,且MDSCs高表达CCR5。细胞迁移结果显示,CCR5抑制剂能够抑制TCM对MDSCs的募集作用。小动物活体成像结果显示,MDSCs经CCR5抑制剂处理后其在体内向脾和肝迁移的作用也减弱。结论在小鼠肝癌微环境中CCR5介导MDSCs在肿瘤微环境中迁移,抑制MDSCs的CCR5可能为肝癌的临床治疗提供新思路。
Objective To explore the role of CCR5 gene in the migration of myeloid-derived suppressor cells(MDSCs) into the mouse liver cancer microenvironment. Methods A mouse model of orthotopic liver cancer was established and changes in the MDSCs were detected by flow cytometry. To observe the effect of mouse liver cancer cell H22(T-cell) conditional medium(TCM) on the migration of MDSCs, these cells were sorted and pretreated with 100 nmol/L CCR5 inhibitor. Finally, DiR-labeled MDSCs were injected into the tail vein of mice with in situ hepatocarcinoma to observe the effect of the CCR5 inhibitor on the migration of MDSCs in vivo. Results The expression of MDSCs in bone marrow, spleen, and liver was significantly higher in the mouse model of orthotopic liver cancer than that in normal mice. The purity of MDSCs was 94.5%, and MDSCs expressed CCR5. Cell migration result showed that CCR5 inhibitor reduced the recruitment of MDSCs by TCM. The in vivo imaging analysis showed that the ability of MDSCs to migrate to the spleen and liver in vivo was weakened after treatment with CCR5 inhibitor. Conclusions CCR5-mediated MDSCs accumulate in the mouse liver cancer microenvironment. Inhibition of CCR5 in MDSCs may provide a new idea for cancer treatment.
Objective To explore the role of CCR5 gene in the migration of myeloid-derived suppressor cells(MDSCs) into the mouse liver cancer microenvironment. Methods A mouse model of orthotopic liver cancer was established and changes in the MDSCs were detected by flow cytometry. To observe the effect of mouse liver cancer cell H22(T-cell) conditional medium(TCM) on the migration of MDSCs, these cells were sorted and pretreated with 100 nmol/L CCR5 inhibitor. Finally, DiR-labeled MDSCs were injected into the tail vein of mice with in situ hepatocarcinoma to observe the effect of the CCR5 inhibitor on the migration of MDSCs in vivo. Results The expression of MDSCs in bone marrow, spleen, and liver was significantly higher in the mouse model of orthotopic liver cancer than that in normal mice. The purity of MDSCs was 94.5%, and MDSCs expressed CCR5. Cell migration result showed that CCR5 inhibitor reduced the recruitment of MDSCs by TCM. The in vivo imaging analysis showed that the ability of MDSCs to migrate to the spleen and liver in vivo was weakened after treatment with CCR5 inhibitor. Conclusions CCR5-mediated MDSCs accumulate in the mouse liver cancer microenvironment. Inhibition of CCR5 in MDSCs may provide a new idea for cancer treatment.
引文
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[22] Pervaiz A,Ansari S,Berger MR,et al.CCR5 blockage by maraviroc induces cytotoxic and apoptotic effects in colorectal cancer cells [J].Med Oncol,2015,32:158.
[23] Mencarelli A,Graziosi L,Renga B,et al.CCR5 antagonism by maraviroc reduces the potential for gastri ccancer cell dissemination [J].Transl Oncol,2013,6:784-793.
[24] Velasco-Velazquez M,Jiao X,De La Fuente M,et al.CCR5 antagonist blocks metastasis of basal breast cancer cells [J].Cancer Res,2012,72:3839-3850.
[25] Sicoli D,Jiao X,Ju X,et al.CCR5 receptor antagonists block metastasis to bone of v-Src oncogene-transformed metastatic prostate cancer cell lines [J].Cancer Res,2014,74:7103-7114.
[26] Tang Q,Jiang J,Liu J.CCR5 blockade suppresses melanoma development through inhibition of IL-6-Stat3 pathway via upregulation of SOCS3 [J].In?ammation,2015,38:2049-2056.
[27] Zhang Y,Lv D,Kim HJ,et al.A novel role of hematopoietic CCL5 in promoting triple-negative mammary tumor progression by regulating generation of myeloid-derived suppressor cells [J].Cell Res,2013,23:394-408.
[2] Sica A,Massarotti M.Myeloid suppressor cells in cancer and autoimmunity [J].J Autoimmun,2017,85:117-125.
[3] Gabrilovich DI,Ostrand-Rosenberg S,Bronte V.Coordinated regulation of myeloid cells by tumours [J].Nat Rev Immunol,2012,12:253-268.
[4] 高苗苗,张纪元,邹正升.髓系来源的抑制细胞在肝病发病中的作用研究进展 [J].实用肝脏病杂志,2018,21(5):813-817.
[5] Yin Z,Li C,Wang J,Xue L.Myeloid-derived suppressor cells:Roles in the tumor microenvironment and tumor radiotherapy [J].Int J Cancer,2019,144(5):933-946.
[6] Elwan N,Salem ML,Kobtan A,et al.High numbers of myeloid derived suppressor cells in peripheral blood and ascitic fluid of cirrhotic and HCC patients [J].Immunol Invest,2018,47(2):169-180.
[7] Highfill SL,Cui Y,Giles AJ,et al.Disruption of CXCR2-mediated MDSC tumor trafficking enhances anti-PD1 efficacy [J].Sci Transl Med,2014,6:237ra67.
[8] Katoh H,Wang D,Daikoku T,et al.CXCR2-expressing myeloid-derived suppressor cells are essential to promote colitis-associated tumorigenesis [J].Cancer Cell,2013,24:631-644.
[9] Sawanobori Y,Ueha S,Kurachi M,et al.Chemokine-mediated rapid turnover of myeloid-derived suppressor cells in tumor-bearing mice [J].Blood,2008,111:5457-5466.
[10] Combadiere C,Ahuja SK,Tiffany HL,Murphy PM.Cloning and functional expression of CC CKR5,a human monocyte CC chemokine receptor selective for MIP-1α,MIP-1β,and RANTES [J].J Leukoc Biol,1996,60(1):147-152.
[11] Cambien B,Richard-Fiardo P,Karimdjee BF,et al.CCL5 neutralization restricts cancer growth and potentiates the targeting of PDGFRbeta in colorectal carcinoma [J].PLoS One,2011,6:e28842.
[12] Sicoli D,Jiao X,Ju X,et al.CCR5 receptor antagonists block metastasis to bone of v-Src oncogene-transformed metastatic prostate cancer cell lines [J].Cancer Res,2014,74:7103-7114.
[13] Velasco-Velazquez M,Xolalpa W,Pestell RG.The potential to target CCL5/CCR5 in breast cancer [J].Expert Opin Ther Targets,2014,18:1265-1275.
[14] Halama N,Zoernig I,Berthel A,et al.Tumoral immune cell exploitation in colorectal cancer metastases can be targeted effectively by anti-CCR5 therapy in cancer patients [J].Cancer Cell,2016,29:587-601.
[15] Groth C,Hu X,Weber R,et al.Immunosuppression mediated by myeloid-derived suppressor cells (MDSCs) during tumour progression [J].Br J Cancer,2019,120:16-25.
[16] Appay V,Rowland-Jones SL.RANTES:a versatile and controversial chemokine [J].Trends Immunol,2001,22:83-87.
[17] Zhao W,Xu Y,Xu J,et al.Subsets of myeloid-derived suppressor cells in hepatocellular carcinoma express chemokines and chemokine receptors differentially [J].Int Immunopharmacol,2015,26:314-321.
[18] 赵文秀,张正奇,许雅苹,等.不同流式抗体分选小鼠原位肝癌模型中髓系来源抑制性细胞的比较 [J].中国实验动物学报,2013,21(4):42-46.
[19] Xu Y,Zhao W,Wu D,et al.Isolation of myeloid-derived suppressor cells subsets from spleens of orthotopic liver cancer-bearing mice by fluorescent-activated and magnetic-activated cell sorting:similarities and differences [J].Int J Clin Exp Pathol,2014,7:7545-7553.
[20] Chang AL,Miska J,Wainwright DA,et al.CCL2 produced by the glioma microenvironment is essential for the recruitment of regulatory T cells and myeloid-derived suppressor cells [J].Cancer Res,2016,76:5671-5682.
[21] Schlecker E,Stojanovic A,Eisen C,et al.Tumor-in?ltrating monocytic myeloid-derived suppressor cells mediate CCR5-dependent recruitment of regulatory Tcells favoring tumor growth [J].J Immunol,2012,189:5602-5611.
[22] Pervaiz A,Ansari S,Berger MR,et al.CCR5 blockage by maraviroc induces cytotoxic and apoptotic effects in colorectal cancer cells [J].Med Oncol,2015,32:158.
[23] Mencarelli A,Graziosi L,Renga B,et al.CCR5 antagonism by maraviroc reduces the potential for gastri ccancer cell dissemination [J].Transl Oncol,2013,6:784-793.
[24] Velasco-Velazquez M,Jiao X,De La Fuente M,et al.CCR5 antagonist blocks metastasis of basal breast cancer cells [J].Cancer Res,2012,72:3839-3850.
[25] Sicoli D,Jiao X,Ju X,et al.CCR5 receptor antagonists block metastasis to bone of v-Src oncogene-transformed metastatic prostate cancer cell lines [J].Cancer Res,2014,74:7103-7114.
[26] Tang Q,Jiang J,Liu J.CCR5 blockade suppresses melanoma development through inhibition of IL-6-Stat3 pathway via upregulation of SOCS3 [J].In?ammation,2015,38:2049-2056.
[27] Zhang Y,Lv D,Kim HJ,et al.A novel role of hematopoietic CCL5 in promoting triple-negative mammary tumor progression by regulating generation of myeloid-derived suppressor cells [J].Cell Res,2013,23:394-408.