摘要
目的探讨基质细胞衍生因子-1(SDF-1)对口腔鳞状细胞癌肿瘤相关巨噬细胞(TAMs)亚型的影响及其制。方法将对数生长期口腔鳞状细胞癌MOC-2细胞随机分为SDF-1诱导组和Cilengitide抑制组,SDF-1诱导组细胞给予100 mg·L^-1 SDF-1100μL诱导24 h,Cilengitide抑制组细胞给予25 mg·L^-1 Cilengitide 100μL抑制1 h后再加入100 mg·L^-1 SDF-1100μL诱导24 h。将C57BL/6J小鼠随机分为SDF-1诱导组(n=12)和Cilengitide抑制组(n=12),SDF-1诱导组小鼠经后背皮下注射50μL基质胶和50μL SDF-1组MOC-2细胞,Cilengitide抑制组小鼠经后背皮下注射50μL基质胶和50μL Cilengitide抑制组MOC-2细胞;4 d后隔日测量1次肿瘤体积,绘制小鼠肿瘤负荷曲线。待肿瘤体积>1500 mm 3时处死小鼠,取肿瘤组织制备单细胞悬液,采用磁性活化细胞分选联合流式细胞术检测TAMs亚型改变,免疫组织化学染色法检测TAMs中CD11b及TAMs各亚型中CD206的表达。结果2组小鼠的肿瘤负荷曲线显示,SDF-1诱导组小鼠肿瘤生长速度较Cilengitide抑制组快。SDF-1诱导组小鼠肿瘤组织中CD45阳性细胞占比显著高于Cilengitide抑制组(t=2.420,P<0.05);SDF-1诱导组小鼠肿瘤组织中CD11b阳性细胞占比显著高于Cilengitide抑制组(t=2.571,P<0.05)。SDF-1诱导组小鼠肿瘤组织中CD45阳性细胞中M2亚型细胞占比显著高于Cilengitide抑制组(t=2.810,P<0.05),M1亚型细胞占比显著低于Cilengitide抑制组(t=2.620,P<0.05)。SDF-1诱导组和Cilengitide抑制组小鼠肿瘤组织中央区CD206、组织相容性复合体Ⅱ(MHCⅡ)表达评分均显著低于外周区(P<0.05);Cilengitide抑制组小鼠肿瘤组织中央区CD206、MHCⅡ表达评分显著低于SDF-1诱导组(P<0.05)。结论SDF-1可能是通过CXC家族趋化因子受体4(CXCR4)-整合蛋白ανβ3(ITGανβ3)生物轴诱导口腔鳞状细胞癌TAMs亚型改变,干扰CXCR4-ITGανβ3的相互作用有可能成为口腔鳞状细胞癌免疫治疗的新方法。
Objective To investigate the effect of stromal cell derived factor-1(SDF-1)on the tumor-associated macrophages(TAMs)subtype in oral squamous cell carcinoma(OSCC).Methods The OSSC MOC-2 cells in logarithmic growth phase were taken and randomly divided into the SDF-1 induction group and Cilengitide inhibition group.The cells in the SDF-1 induction group were treated with 100 mg·L^-1 SDF-1100μL for 24 h;the cells in the Cilengitide inhibition group were treated with 25 mg·L^-1 Cilengitide 100μL for 1 h,and then they were treated with 100 mg·L^-1 SDF-1100μL for 24 h.C57BL/6J mice were randomly divided into the SDF-1 induction group(n=12)and Cilengitide inhibition group(n=12).The mice in the SDF-1 induction group were subcutaneously injected with 50μL matrigel and 50μL MOC-2 cells of the SDF-1 induction group;the mice in Cilengitide inhibition group were subcutaneously injected with 50μL matrigel and 50μL MOC-2 cells of the Cilengitide inhibition group;after 4 days,the tumor volumes of mice in the two groups were measured every other day and the tumor load curve were drawn.When the tumor volume was more than 1500 mm 3,the mice were killed and the tumor tissues were taken to prepare single cell suspension.The TAMs subtypes were detected by magnetic activated cell sorting and flow cytometry;the expression of CD11b in TAMs and the expression of CD206 in TAMs subtypes were detected by immunohisto chemistry staining.Results The tumor load curve of mice in the two groups showed that the tumor growth rate of mice in the SDF-1 induction group was faster than that in the Cilengitide inhibition group.The percentage of CD45 positive cells in tumor tissue of mice in the SDF-1 induction group was significantly higher than that in the Cilengitide inhibition group(t=2.420,P<0.05);the proportion of CD11b positive cells in tumor tissue of mice in the SDF-1 induction group was significantly higher than that in the Cilengitide inhibition group(t=2.571,P<0.05).The proportion of M2 subtype cells in CD45 positive cells in tumor tissue of mice in the SDF-1 induction group was significantly higher than that in the Cilengitide inhibition group(t=2.810,P<0.05),and the proportion of M1 subtype cells was significantly lower than that in the Cilengitide inhibition group(t=2.620,P<0.05).The expression scores of CD206 and major histocompatibility complexⅡ(MHCⅡ)in the central region of the tumor tissue were significantly lower than those in the peripheral region in the SDF-1 induction group and the Cilengitide inhibition group(P<0.05);the expression scores of CD206 and MHCⅡin the central region of the tumor tissue of mice in the Cilengitide inhibition group were significantly lower than those in the SDF-1 induction group(P<0.05).Conclusion SDF-1 may induce TAMs phenotype changes of OSCC through the C-X-C chemokine receptor 4(CXCR4)-integrinανβ3(ITGανβ3)biological axis,and interfering with the interaction of CXCR4-ITGανβ3 may become a new immunotherapy method for OSCC.
作者
李鹏
孙丽艳
郭兰伟
李文鹿
罗瑞华
齐金星
LI Peng;SUN Liyan;GUO Lanwei;LI Wenlu;LUO Ruihua;QI Jinxing(Department of Head Neck and Thyroid Surgery,the Affiliated Cancer Hospital of Zhengzhou University,Zhengzhou 450008,Henan Province,China;Department of Pediatric Dentistry,SATH Stomatology Hospital of Henan University,Zhengzhou 450008,Henan Province,China;Department of Cancer Control and Research,the Affiliated Cancer Hospital of Zhengzhou University,Zhengzhou 450008,Henan Province,China;Department of Oral and Maxillofacial Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan Province,China)
出处
《新乡医学院学报》
CAS
2020年第12期1108-1113,共6页
Journal of Xinxiang Medical University
基金
国家自然科学基金项目(编号:81703298,81402578)
河南省医学科技攻关计划项目(编号:2018020488)
河南省重点研发与推广专项(编号:192102310354)。
