不同来源细胞因子诱导杀伤细胞体外抗肿瘤作用比较探究.docVIP

不同来源细胞因子诱导杀伤细胞体外抗肿瘤作用比较探究 　　[摘要] 目的 比较来自脐带血和恶性肿瘤患者外周血的细胞因子诱导的杀伤细胞（cytokine-induced killer，CIK）体外抗肿瘤作用的差异。 方法 体外诱导培养肺癌患者外周血和脐血来源的CIK细胞，MTT法和流式细胞术检测诱导的CIK细胞的抗肿瘤活性及机制。 结果 来自肺癌患者外周血和脐血的CIK细胞中CD3+细胞、CD3+CD56+ 和CD3+CD8+细胞百分比均伴随培养时间的延长而升高，组间比较无统计学差异；诱导培养21 d时，脐血CIK细胞和肺癌患者外周血来源的CIK细胞的增殖率分别为（185.76plusmn;39.68）%、（254.32plusmn;74.60）%，二者之间差异有统计学意义（P=0.0012）；脐血CIK细胞的抗肿瘤活性高于肺癌患者外周血来源的CIK细胞（效靶比10∶1，靶细胞为K562细胞时，P=0.0016；靶细胞为A549细胞时，P=0.0022）；脐血CIK细胞CD107a表达高于来自肺癌患者外周血的CIK细胞 （靶细胞为K562细胞时，P=0.0469；靶细胞为A549细胞时，P=0.0413）；脐血CIK细胞IFN-gamma;和TNF-alpha;的分泌水平高于肺癌患者CIK细胞（P=0.0352；P=0.0389）；脐血CIK细胞和肺癌患者CIK细胞诱导K562细胞凋亡的百分率为（25.30plusmn;4.87）%和（19.87plusmn;5.41）%，二者之间差异无统计学意义（P=0.2658）。 结论 流式细胞术可同时检测CIK细胞的表型、功能及作用机制。本研究为临床应用CIK治疗提供更多选择和实验室依据。　　[关键词] 细胞因子诱导的杀伤细胞；流式细胞术；免疫治疗；CD107a　　[中图分类号] R392.9；R730.2 [文献标识码] A [文章编号] 1673-9701（2016）19-0038-05　　[Abstract] Objective To compare difference of the anti-tumor response of CIK cells from peripheral blood of patients with lung cancer and umbilical cord blood in vitro. Methods Peripheral monocytes were isolated from patients with lung cancer， and cord blood monocytes from healthy donors. Isolated cells were cultured and induced as CIK cells in vitro. The anti-tumor activities were measured by MTT assay and flow cytometry. Intracellular staining was used to test the secretion of cytokines. Results The percentage of CD3+， CD3+CD56+ and CD3+CD8+ cells in cord blood CIK and peripheral blood CIK of patients with lung cancer was increased with the prolongation of culture time， and there was no significant difference between the two groups； After cultured for 21 days， the proliferation rate of cord blood CIK cells and lung cancer patients’ peripheral blood CIK cells was（185.76plusmn;39.68）% and（254.32plusmn;74.60）%， respectively， and there was significant difference between the two group （P=0.0012）； The anti-tumor activity of cord blood CIK cells was higher than that of lung cancer patients’ peripheral blood CIK cells（effector-target ratio of 10：1， when the