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目的:分析细胞因子诱导的杀伤细胞(cytokine induced killer cell,CIK)抗肿瘤活性增强的分子机制。方法:从健康人志愿者和恶性肿瘤患者PBMC培养获得CIK细胞,用LDH释放法分别对PBMC和CIK进行体外杀伤肿瘤细胞的活性测定,用流式细胞术分析PBMC和CIK细胞群中颗粒溶素(granulysin,GNLY)和穿孔素(perforin,PFN)的表达。结果:在效靶比为5∶1时,健康人和肿瘤患者PBMC对肿瘤细胞K562的裂解率分别为6.33%和1.29%,两组CIK细胞对肿瘤细胞K562的裂解率分别为14.7%和14.16%,PB-MC与CIK对细胞的裂解率显著提高,P<0.01;GNLY阳性表达细胞百分比由4.43%和1.95%上升到19.15%和17.52%,P<0.01。但PFN细胞却由21.91%和14.69%降低到4.96%和3.7%,P<0.05。结论:肿瘤患者PBMC抗肿瘤活性下降可能与该细胞中GN-LY和PFN表达减少有关;CIK细胞杀伤肿瘤活性的提高可能与GNLY的表达增加相关;PFN表达下降可能是维持CIK细胞在体外大量增殖的调节因素。
Objective: To analyze the molecular mechanism of antitumor activity enhanced by cytokine induced killer cell (CIK). Methods: CIK cells were obtained from PBMC of healthy volunteers and malignant tumor patients. The cytotoxic activity of PBMC and CIK in vitro were assayed by LDH release assay. The levels of granulysin in PBMC and CIK cell population were analyzed by flow cytometry (granulysin, GNLY) and perforin (PFN) expression. Results: When the ratio of target to target was 5: 1, the rates of cleavage of K562 in PBMC of healthy and cancer patients were 6.33% and 1.29%, respectively. The cleavage rates of CIK cells to K562 were 14.7% and 14.16 %, The rate of cell lysis was significantly increased by PB-MC and CIK, P <0.01; the percentage of GNLY positive cells increased from 4.43% and 1.95% to 19.15% and 17.52% respectively, P <0.01. However, PFN cells decreased from 21.91% and 14.69% to 4.96% and 3.7% respectively, P <0.05. CONCLUSION: The decrease of anti-tumor activity of PBMC in tumor patients may be related to the decrease of GN-LY and PFN expression. The increase of cytotoxicity of CIK cells may be related to the increase of GNLY expression. The decrease of PFN expression may be the reason of CIK cell proliferation in vitro The adjustment factor.