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目的:探讨特异性针对表皮生长因子受体变异体Ⅲ(epidermal growth factor receptor variantⅢ,EGFRvⅢ)的单克隆抗体CH12(CH12单抗)对EGFRvⅢ阳性表达的肺鳞癌生长的影响。方法:采用慢病毒感染的方法建立EGFRvⅢ过表达的人肺鳞癌SK-MES-1-EGFRvⅢ和NCI-H520-EGFRvⅢ细胞。FCM法检测CH12单抗与SKMES-1-EGFRvⅢ和NCI-H520-EGFRvⅢ细胞的结合能力。检测由CH12单抗诱导的外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)对SK-MES-1-EGFRvⅢ和NCI-H520-EGFRvⅢ细胞的抗体依赖性细胞介导的细胞毒性作用(antibody-dependent cell-mediated cytotoxicity,ADCC)。裸鼠皮下接种SK-MES-1-EGFRvⅢ细胞,建立EGFRvⅢ过表达的肺鳞癌裸鼠移植瘤模型。实验分为3组:(1)腹腔注射CH12单抗组;(2)腹腔注射PBS组(阴性对照);(3)腹腔注射人鼠嵌合性C225单克隆抗体(阳性对照);观察裸鼠体内肿瘤的生长情况,并绘制生长曲线。结果:成功构建了EGFRvⅢ过表达的肺鳞癌SK-MES-1-EGFRvⅢ和NCI-H520-EGFRvⅢ细胞;FCM法检测结果显示,CH12单抗能够与SK-MES-1-EGFRvⅢ及H520-EGFRvⅢ细胞高度结合,而与亲本SKMES-1及NCI-H520细胞不结合。ADCC结果表明,CH12单抗能够介导PBMCs对SK-MES-1-EGFRvⅢ及H520-EGFRvⅢ细胞的细胞毒杀伤作用(P值均<0.01)。对裸鼠体内移植瘤的治疗结果显示,CH12单抗能够完全清除裸鼠体内的SK-MES-1-EGFRvⅢ细胞移植瘤。结论:体外和体内研究均证实,CH12单抗对EGFRvⅢ过表达的肺鳞癌细胞的生长具有明显的抑制作用。
OBJECTIVE: To investigate the effect of monoclonal antibody CH12 (CH12 monoclonal antibody) targeting epidermal growth factor receptor variant Ⅲ (EGFRvⅢ) on the growth of lung squamous cell carcinoma (EGFRvⅢ) -positive lung squamous cell carcinoma. Methods: Human lung squamous cell carcinoma SK-MES-1-EGFRvⅢ and NCI-H520-EGFRvⅢ overexpressed by EGFRvIII were constructed by lentivirus infection. The binding ability of CH12 monoclonal antibody to SKMES-1-EGFRvIII and NCI-H520-EGFRvIII cells was detected by FCM. Antibody-dependent cell-mediated cytotoxicity of SK-MES-1-EGFRvIII and NCI-H520-EGFRvIII cells induced by CH12 monoclonal antibody (PBMCs) cell-mediated cytotoxicity, ADCC). The nude mice were inoculated with SK-MES-1-EGFRvIII cells subcutaneously to establish a model of lung squamous cell carcinoma xenograft in which EGFRvIII was overexpressed. The experiment was divided into three groups: (1) intraperitoneal injection of CH12 monoclonal antibody; (2) intraperitoneal injection of PBS group (negative control); (3) intraperitoneal injection of human chimeric C225 monoclonal antibody (positive control) In vivo tumor growth, and draw the growth curve. Results: The SK-MES-1-EGFRvⅢ and NCI-H520-EGFRvⅢ cells were successfully constructed with EGFRvⅢ overexpression. The results of FCM showed that CH12 McAb was able to bind with SK-MES-1-EGFRvⅢ and H520- Highly bound, but not with parental SKMES-1 and NCI-H520 cells. ADCC results showed that the monoclonal antibody of CH12 could mediate the cytotoxicity of PBMCs on SK-MES-1-EGFRvIII and H520-EGFRvIII cells (all P <0.01). The results of the treatment of transplanted tumors in nude mice showed that CH12 monoclonal antibody can completely remove the SK-MES-1-EGFRvIII cell xenografts in nude mice. CONCLUSION: In vitro and in vivo studies have confirmed that CH12 monoclonal antibody has a significant inhibitory effect on the growth of lung squamous cell carcinoma cells overexpressing EGFRvIII.