二甲双胍介导的增强CD8阳性T细胞肿瘤浸润与增加的肿瘤灌注的相关性
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摘要
目的探讨二甲双胍产生的增强CD8阳性T细胞肿瘤浸润作用与增加的肿瘤灌注之间的相关性。方法 6～8周雌性小鼠乳腺脂肪垫接种4T1细胞后9d,随机分为对照组及二甲双胍干预组。提取标本前15min,经尾静脉注射TRITC标记的Lectin溶液,提取标本并制备成冰冻切片后行CD8抗体荧光染色,用激光共聚焦显微镜采集结果。用Spearman分析方法观察相关性。结果二甲双胍干预组的CD8阳性细胞浸润数量[(120.8±26.9)个/mm~2 vs.(67.9±22.9)个/mm~2]及Lectin阳性信号面积[(18 357.0±5 865.0)像素/μm~2 vs.(10 233.0±4 434.0)像素/μm~2]显著高于对照组(P<0.05)。两组的CD8阳性T细胞浸润数量与Lectin阳性信号面积均呈线性相关(对照组:R=0.884,P=0.007;二甲双胍组:R=0.928,P<0.001)。结论增加的肿瘤灌注与二甲双胍介导的增强CD8阳性T细胞肿瘤浸润密切相关。
Objective To investigate the linear correlation between metformin-induced increase in the number of CD8~+T cells infiltrated into the cancer and metformin-mediated elevation of tumor blood perfusion.Methods4 T1 murine breast cancer cells were orthotopically injected into the fat pad of the 4th breast of BALB/C mice.Nine days later,the mice were randomly divided into the control and metformin groups.Fifteen minutes before extraction,TRITC-conjugated lectin was injected into the tail vein of the mice.Then,the extracted tumor tissues were prepared for frozen sections,which were then stained with anti-CD8 antibody for further immunofluorescent detection.Spearman correlation analysis was performed to observe the correlation between the parameters.ResultsBoth the infiltration of CD8~+T cells [(120.8±26.9)/mm~2 vs.(67.9±22.9)/mm~2]and lectin-perfused area[(18 357.0±5 865.0)pixels/μm~2 vs.(10 233.0±4 434.0)pixels/μm~2]were significantly higher in metformin-treated group than in the untreated group.In both groups,the number of the infiltrated CD8~+T cells was linearly correlated with lectin-perfused area(control:R =0.884,P=0.007;metformin:R =0.928,P<0.001).ConclusionIncreased tumor perfusion is closely correlated with metformin-induced increase in the infiltration of CD8~+T cells into the tumor.
Objective To investigate the linear correlation between metformin-induced increase in the number of CD8~+T cells infiltrated into the cancer and metformin-mediated elevation of tumor blood perfusion.Methods4 T1 murine breast cancer cells were orthotopically injected into the fat pad of the 4th breast of BALB/C mice.Nine days later,the mice were randomly divided into the control and metformin groups.Fifteen minutes before extraction,TRITC-conjugated lectin was injected into the tail vein of the mice.Then,the extracted tumor tissues were prepared for frozen sections,which were then stained with anti-CD8 antibody for further immunofluorescent detection.Spearman correlation analysis was performed to observe the correlation between the parameters.ResultsBoth the infiltration of CD8~+T cells [(120.8±26.9)/mm~2 vs.(67.9±22.9)/mm~2]and lectin-perfused area[(18 357.0±5 865.0)pixels/μm~2 vs.(10 233.0±4 434.0)pixels/μm~2]were significantly higher in metformin-treated group than in the untreated group.In both groups,the number of the infiltrated CD8~+T cells was linearly correlated with lectin-perfused area(control:R =0.884,P=0.007;metformin:R =0.928,P<0.001).ConclusionIncreased tumor perfusion is closely correlated with metformin-induced increase in the infiltration of CD8~+T cells into the tumor.
引文
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[5]SCHOTT S,BIERHAUS A,SCHUETZ F,et al.Therapeutic effects of metformin in breast cancer:Involvement of the immune system?[J].Cancer Immunol,Immunother,2011,60(9):1221-1225.
[6]EIKAWA S,NISHIDA M,MIZUKAMI S,et al.Immune-mediated antitumor effect by type 2 diabetes drug,metformin[J].Proc Nati Acad Sci,2015,112(6):1809-1814.
[7]KARYAMPUDI L,LAMICHHANE P,SCHEID AD,et al.Accumulation of memory precursor CD8 T cells in regressing tumors following combination therapy with vaccine and antiPD-1 antibody[J].Cancer Res,2014,74(11):2974-2985.
[8]RANKIN EB,GIACCIA AJ.The role of hypoxia-inducible factors in tumorigenesis[J].Cell Death Differ,2008,15(4):678-685.
[9]KUCHNIO A,MOENS S,BRUNING U,et al.The cancer cell oxygen sensor PHD2 promotes metastasis via activation of cancer-associated fibroblasts[J].Cell Reports,2015,12(6):992-1005.
[10]KUNZ M,IBRAHIM SM.Molecular responses to hypoxia in tumor cells[J].Mol Cancer,2003,2(1):23.
[11]WANG J,LI G,WANG Y,et al.Suppression of tumor angiogenesis by metformin treatment via a mechanism linked to targeting of HER2/HIF-1alpha/VEGF secretion axis[J].Oncotarget,2015,6(42):44579-44592.
[12]HUANG Y,GOEL S,DUDA DG,et al.Vascular normalization as an emerging strategy to enhance cancer immunotherapy[J].Cancer Res,2013,73(10):2943-2948.
[13]CARRETERO R,SEKTIOGLU IM,GARBI N,et al.Eosinophils orchestrate cancer rejection by normalizing tumor vessels and enhancing infiltration of CD8+T cells[J].Nat Immunol,2015,16(6):609.
[14]WANG JC,LI GY,LI PP,et al.Suppression of hypoxia-induced excessive angiogenesis by metformin via elevating tumor blood perfusion[J].Oncotarget,2017,8(43):73892-73904.