下咽鳞状细胞癌CAR-EGFR修饰T细胞构建的初步研究

Preliminary study on the construction of CAR-EGFR modified T cells(EGFR-CART) in hypopharyngeal squamous cell carcinoma

目的构建具有靶向功能下咽鳞状细胞癌Fadu细胞系,EGFR-CART细胞并进行验证,为CART细胞技术在下咽鳞状细胞癌治疗应用临床前基础进行探索。方法通过慢病毒转染构建EGFR-CART细胞及CART-对照细胞,FACS荧光检测CART细胞上EGFR抗体的表达,验证Fadu细胞系及HCT-116细胞系中EGFR的表达。结果成功构建了EGFR-CART细胞及CART-对照细胞,感染效率约为67%,二者在细胞亚群的分布上没有明显区别。靶细胞功能验证发现HCT-116中EGFR低表达,Fadu中EGFR高表达,确定HCT-116作为EGFR-CART的非靶细胞,Fadu作为靶细胞。结论本研究成功制备并验证了具有靶向功能的下咽鳞状细胞癌Fadu细胞系的EGFR-CART细胞,为CART技术在下咽癌治疗中的应用打下了前期基础。

OBJECTIVE Construct and validate the EGFR-CART （Epidermal growth factor receptorchimeric antigen receptor T） cells targeting the Fadu cell line of hypopharyngeal squamous cell carcinoma, and provide the preclinical basis for the application of CART cell technology in hypopharyngeal squamous cell carcinoma. METHODS EGFR-CART cells and CART-control cells were constructed by lentivirus transfection. The expression of EGFR antibody in CART cells was detected by FACS fluorescence detection, and the expression of EGFR in Fadu cell line and HCT-116 cell line was verified. RESULTS EGFR-CART cells and CART-control cells were successfully constructed, the infection efficiency was about 67%, and there was no significant difference between the two groups in the distribution of cell subsets. HCT-116 is selected as the non target cells of EGFR-CART, Fadu as target cell. CONCLUSION In this study, we successfully prepared and validated the EGFR-CART cells targeting the Fadu cell line of hypopharyngeal squamous cell carcinoma, and laid the foundation for the application of CART technique in the treatment of hypopharyngeal carcinoma.