不可逆电穿孔消融术联合PD-1抑制剂治疗小鼠肝癌效果初步研究
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摘要
目的探讨不可逆电穿孔(IRE)消融术联合程序性细胞死亡蛋白(PD)-1抑制剂治疗小鼠肝癌效果及肿瘤微环境变化。方法构建小鼠皮下肝癌模型,随机分为对照组、IRE组、PD-1组和IRE联合PD-1组。采用实时荧光定量聚合酶链反应(PCR)和免疫组化分析不同组别不同时间点肿瘤组织CD8和Foxp3细胞表达,流式细胞学检测外周血T细胞分群,并描绘肿瘤生长曲线。结果 IRE消融术联合PD-1抑制剂治疗后,小鼠肿瘤体积明显缩小,外周血和肿瘤组织CD8+T细胞表达均增多,调节性T细胞(Treg)水平明显下降。结论 IRE消融术联合PD-1抑制剂能有效减少肿瘤负荷,引起肿瘤微环境改变,为肝癌消融联合免疫治疗提供了新的方向。
Objective To investigate the efficacy of irreversible electroporation(IRE) ablation combined with programmed cell death protein-1(PD-1) inhibitor in the treatment of hepatocellular carcinoma(HCC) in experimental mice, and to discuss the changes in tumor microenvironment. Methods The subcutaneous HCC model was established in mice. The mouse models were randomly divided into the following four groups: control group, IRE group, PD-1 inhibitor group and IRE combined with PD-1 inhibitor group(combination group). The real-time fluorescence quantitative polymerase chain reaction(RTq PCR) and immunohistochemistry were used to determine the expressions of CD8+T cell and regulatory T cell(Treg) in tumor tissue of the mice of all groups at different point of time. By using flow cytometry, the T cell subgroups in peripheral blood were tested, and the growth curves of tumor were drawn. Results In combination group, after receiving IRE combined with PD-1 inhibitor therapy, the tumor volume of the mice was obviously reduced, the expressions of CD8+T cell in peripheral blood and tumor tissue were increased and the Treg levels were significantly decreased. Conclusion In treating HCC of experimental mice, the combination use of IRE and PD-1 inhibitor can effectively reduce the tumor load and induce the changes of tumor microenvironment, which provides a new direction for ablation and immunotherapy of HCC.
Objective To investigate the efficacy of irreversible electroporation(IRE) ablation combined with programmed cell death protein-1(PD-1) inhibitor in the treatment of hepatocellular carcinoma(HCC) in experimental mice, and to discuss the changes in tumor microenvironment. Methods The subcutaneous HCC model was established in mice. The mouse models were randomly divided into the following four groups: control group, IRE group, PD-1 inhibitor group and IRE combined with PD-1 inhibitor group(combination group). The real-time fluorescence quantitative polymerase chain reaction(RTq PCR) and immunohistochemistry were used to determine the expressions of CD8+T cell and regulatory T cell(Treg) in tumor tissue of the mice of all groups at different point of time. By using flow cytometry, the T cell subgroups in peripheral blood were tested, and the growth curves of tumor were drawn. Results In combination group, after receiving IRE combined with PD-1 inhibitor therapy, the tumor volume of the mice was obviously reduced, the expressions of CD8+T cell in peripheral blood and tumor tissue were increased and the Treg levels were significantly decreased. Conclusion In treating HCC of experimental mice, the combination use of IRE and PD-1 inhibitor can effectively reduce the tumor load and induce the changes of tumor microenvironment, which provides a new direction for ablation and immunotherapy of HCC.
引文
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[13]Zou W,Wolchok JD,Chen L.PD-L1(B7-H1)and PD-1 pathway blockade for cancer therapy:mechanisms,response biomarkers,and combinations[J].Sci Transl Med,2016,8:328rv4.
[14]Inoue H,Tani K.Multimodal immunogenic cancer cell death as a consequence of anticancer cytotoxic treatments[J].Cell Death Differ,2014,21:39-49.
[15]徐晓林,朱争艳,经翔.肝细胞癌患者射频消融前后的免疫状态研究进展[J].介入放射学杂志,2017,26:87-91.
[16]韩宗宏,曹建民,史东宏.肿瘤冷冻消融免疫效应研究的进展[J].介入放射学杂志,2013,22:1055-1058.
[17]Rozenblum N,Zeira E,Scaiewicz V,et al.Oncogenesis:an“off-target”effect of radiofrequency ablation[J].Radiology,2015,276:426-432.
[18]Al-Sakere B,Bernat C,Andre F,et al.A study of the immunological response to tumor ablation with irreversible electroporation[J].Technol Cancer Res Treat,2007,6:301-306.
[19]Chen X,Ren Z,Yin S,et al.The local liver ablation with pulsed electric field stimulate systemic immune reaction against hepatocellular carcinoma(HCC)with time-dependent cytokine profile[J].Cytokine,2017,93:44-50.
[2]刘树鹏,秦子淋,陈继冰,等.不可逆电穿孔消融治疗前列腺癌进展[J].介入放射学杂志,2018,27:386-389.
[3]李鑫,刘凤永,袁宏军,等.纳米刀治疗肝癌[J].介入放射学杂志,2017,26:939-943.
[4]Ansari D,Kristoffersson S,Andersson R,et al.The role of irreversible electroporation(IRE)for locally advanced pancreatic cancer:a systematic review of safety and efficacy[J].Scand JGastroenterol,2017,52:1165-1171.
[5]White SB,Zhang Z,Chen J,et al.Early immunologic response of irreversible electroporation versus cryoablation in a rodent model of pancreatic cancer[J].J Vasc Interv Radiol,2018,29:1764-1769.
[6]Bulvik BE,Rozenblum N,Gourevich S,et al.Irreversible electroporation versus radiofrequency ablation:a comparison of local and systemic effects in a small-animal model[J].Radiology,2016,280:413-424.
[7]Distelmaier M,Barabasch A,Heil P,et al.Midterm safety and efficacy of irreversible electroporation of malignant liver tumors located close to major portal or hepatic veins[J].Radiology,2017,285:1023-1031.
[8]El-Khoueiry AB,Sangro B,Yau T,et al.Nivolumab in patients with advanced hepatocellular carcinoma(CheckMate 040):an open-label,non-comparative,phase 1/2 dose escalation and expansion trial[J].Lancet,2017,389:2492-2502.
[9]Shi L,Chen L,Wu C,et al.PD-1 blockade boosts radiofrequency ablation-elicited adaptive immune responses against tumor[J].Clin Cancer Res,2016,22:1173-1184.
[10]Nuccitelli R,Berridge JC,Mallon Z,et al.Nanoelectroablation of murine tumors triggers a CD8-dependent inhibition of secondary tumor growth[J].PLoS One,2015,10:e0134364.
[11]Schachter J,Ribas A,Long GV,et al.Pembrolizumab versus ipilimumab for advanced melanoma:final overall survival results of a multicentre,randomised,open-label phase 3 study(KEYNOTE-006)[J].Lancet,2017,390:1853-1862.
[12]Tumeh PC,Harview CL,Yearley JH,et al.PD-1 blockade induces responses by inhibiting adaptive immune resistance[J].Nature,2014,515:568-571.
[13]Zou W,Wolchok JD,Chen L.PD-L1(B7-H1)and PD-1 pathway blockade for cancer therapy:mechanisms,response biomarkers,and combinations[J].Sci Transl Med,2016,8:328rv4.
[14]Inoue H,Tani K.Multimodal immunogenic cancer cell death as a consequence of anticancer cytotoxic treatments[J].Cell Death Differ,2014,21:39-49.
[15]徐晓林,朱争艳,经翔.肝细胞癌患者射频消融前后的免疫状态研究进展[J].介入放射学杂志,2017,26:87-91.
[16]韩宗宏,曹建民,史东宏.肿瘤冷冻消融免疫效应研究的进展[J].介入放射学杂志,2013,22:1055-1058.
[17]Rozenblum N,Zeira E,Scaiewicz V,et al.Oncogenesis:an“off-target”effect of radiofrequency ablation[J].Radiology,2015,276:426-432.
[18]Al-Sakere B,Bernat C,Andre F,et al.A study of the immunological response to tumor ablation with irreversible electroporation[J].Technol Cancer Res Treat,2007,6:301-306.
[19]Chen X,Ren Z,Yin S,et al.The local liver ablation with pulsed electric field stimulate systemic immune reaction against hepatocellular carcinoma(HCC)with time-dependent cytokine profile[J].Cytokine,2017,93:44-50.