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目的初步验证增强表达CD16分子的CIK细胞联合EGFR单克隆抗体在体外对胃癌细胞的杀伤作用。探讨其治疗胃癌的临床应用前景。方法将CD16基因插入载体pcDNA3.1构建PC-CD16真核表达载体,脂质体瞬时转染CIK细胞增强其CD16表达建立CD16高表达的CIK-CD16细胞系。该细胞系联合EGFR单克隆抗体进行51Cr杀伤实验检测其与胃癌SGC7901细胞结合及杀伤能力并与普通CIK细胞比较。使用流式细胞仪法检测治疗后肿瘤细胞周期与凋亡情况变化。结果51Cr细胞杀伤实验结果提示:CIK-CD16细胞联合EGFR单克隆抗体对胃癌细胞株SGC7901杀伤率显著高于各对照组(P<0.05)。CIK-CD16细胞联合EGFR单克隆抗体处理后肿瘤细胞凋亡率上升。结论初步的体外实验显示,增强CD16表达的CIK细胞联合EGFR单克隆抗体可有效增强免疫效应细胞对胃癌的杀伤作用,具有潜在的临床应用价值。
Objective To verify the killing effect of CIK cells with enhanced expression of CD16 and EGFR monoclonal antibody on gastric cancer cells in vitro. To explore its clinical application of gastric cancer prospects. Methods CD16 gene was inserted into pcDNA3.1 to construct PC-CD16 eukaryotic expression vector. The liposome was transiently transfected into CIK cells to enhance the expression of CD16 and establish the CD16-overexpressing CIK-CD16 cell line. The cell lines combined with EGFR monoclonal antibody 51Cr killing test to detect its binding to gastric cancer SGC7901 cells and cytotoxicity and compared with ordinary CIK cells. The changes of tumor cell cycle and apoptosis after treatment were detected by flow cytometry. Results The results of 51Cr cell killing experiment showed that the killing rate of CIK-CD16 cells combined with EGFR monoclonal antibody on gastric cancer cell line SGC7901 was significantly higher than that of the control group (P <0.05). The apoptosis rate of CIK-CD16 cells treated with EGFR monoclonal antibody increased. Conclusion Preliminary in vitro experiments show that CIK cells enhanced CD16 expression combined with EGFR monoclonal antibody can effectively enhance the killing effect of immune effector cells on gastric cancer and has potential clinical value.