脐带血和乳腺癌患者外周血来源CIK细胞PD-1表达及对MCF-7细胞杀伤

发布时间：2018-11-07 20:25

【摘要】：目的:探讨脐带血和乳腺癌患者外周静脉血来源的CIK细胞程序性死亡分子-l(programmed cell death-1,PD-1)的表达及其对乳腺癌MCF-7细胞的杀伤作用。方法:采集2015年6月至2015年12月在解放军第105医院住院的健康产妇脐带血和乳腺癌患者外周静脉血各5例,分离PBMC,体外培养、扩增CIK细胞。流式细胞术检测不同时间节点两种来源的CIK细胞PD-1表达情况,分别取培养第7、14、21、28天的CIK细胞与MCF-7细胞共培养,细胞计数(CCK-8)法测定CIK细胞对MCF-7细胞的杀伤率,吖啶橙-溴化乙啶双染(AOEB)法观察共培养后CIK细胞凋亡变化,流式细胞术检测CIK细胞凋亡率。结果:随着体外培养时间的延长,脐带血和乳腺癌患者静脉血来源的CIK细胞PD-1表达率均逐渐上升,第14天时脐带血组CIK细胞PD-1表达率低于乳腺癌组[(38.42±4.76)%vs(50.54±3.50)%,P0.05],至第21天后两组PD-1表达率均升高,但差异无统计学意义(P0.05)。培养第7、14、21、28天两组CIK细胞对MCF-7细胞的杀伤率分别为(18.54±3.54)%和(21.74±4.27)%、(71.86±16.86)%和(58.78±24.25)%、(44.32±26.87)%和(43.96±26.04)%、(43.24±24.27)%和(40.28±23.69)%,以培养第14天的脐带血来源的CIK细胞的杀伤活性最强(P0.05)。分析发现,两种来源的CIK细胞PD-1表达水平与CIK细胞的凋亡率呈正相关(r=0.971,r=0.900,均P0.01),与杀伤率呈负相关(r=-0.865,r=-0.885,均P0.01)。结论:活化的CIK细胞表面高表达PD-1,脐带血较乳腺癌患者静脉血来源的CIK细胞表面PD-1水平低;培养第14天的CIK细胞凋亡率均较低,其杀伤能力更强。
[Abstract]:Aim: to investigate the expression of programmed death molecule (l (programmed cell death-1,PD-1) in CIK cells from umbilical cord blood and peripheral venous blood of patients with breast cancer and its killing effect on MCF-7 cells of breast cancer. Methods: from June 2015 to December 2015, 5 cases of healthy maternal umbilical cord blood and 5 cases of peripheral venous blood of breast cancer patients were collected from 105 hospitals of PLA. PBMC, was isolated and cultured in vitro to amplify CIK cells. Flow cytometry was used to detect the expression of PD-1 in CIK cells from two different sources at different time points. The CIK cells were cocultured with MCF-7 cells on day 714, 21 and 28, respectively. The cytotoxicity of CIK cells to MCF-7 cells was determined by cell count (CCK-8) method. The apoptosis of CIK cells after co-culture was observed by acridine orange and ethidium bromide double staining (AOEB) method, and the apoptosis rate of CIK cells was detected by flow cytometry. Results: with the prolongation of culture time in vitro, the PD-1 expression rate of CIK cells from umbilical cord blood and breast cancer patients increased gradually. On the 14th day, the PD-1 expression rate of CIK cells in cord blood group was lower than that in breast cancer group [(38.42 卤4.76)% vs (50.54 卤3.50)%, P0.05]. But the difference was not statistically significant (P0.05). The cytotoxicity of CIK cells to MCF-7 cells was (18.54 卤3.54)% and (21.74 卤4.27)%, (71.86 卤16.86)% and (58.78 卤24.25)%, (44.32 卤26.87)% and (43.96 卤26.04)%, respectively. (43.24 卤24.27)% and (40.28 卤23.69)% respectively. The cytotoxicity of CIK cells from cord blood on the 14th day was the highest (P0.05). It was found that the expression of PD-1 in CIK cells was positively correlated with the apoptosis rate of CIK cells (r = 0.900, all P 0.01), and negatively correlated with the killing rate (r = 0.865, r = 0.885, all P 0.01). Conclusion: the overexpression of PD-1, on the surface of activated CIK cells is lower than that of CIK cells derived from breast cancer, and the apoptotic rate of CIK cells on the 14th day of culture is lower, and the cytotoxicity of CIK cells is stronger.
【作者单位】：解放军第105医院生物治疗中心;安徽医科大学第一附属医院重症医学科;安徽省立医院肿瘤科;
【基金】：南京军区科技创新基金资助项目(No:15MS048)~~
【分类号】：R737.9

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