目的： 研究顺铂、紫杉醇分别对CIK细胞杀伤肺癌A549细胞的影响并初步探讨其可能的分子机制。方法： MTT法分别检测顺铂、紫杉醇处理A549细胞24 h的半数抑制浓度（IC50），LDH释放法检测IC50的顺铂，紫杉醇分别作用24 h对CIK细胞杀伤A549细胞能力的影响；流式细胞术检测顺铂、紫杉醇作用24 h对A549细胞表面NKG2D配体（MICA、MICB、ULBP1、ULBP2、ULBP3）表达的影响，荧光定量PCR检测顺铂、紫杉醇作用对A549细胞内DNA损伤修复基因（ATM、ATR、CHK1、CHK2、P53）表达的影响。结果： 顺铂和紫衫醇对A549细胞作用24 h的IC50分别为70、39 μg/ml。IC50的顺铂、紫杉醇分别作用24 h后，CIK细胞对A549细胞的杀伤活性均明显增强（P<0.05）；同时A549细胞表面MICA、MICB、ULBP2、ULBP3表达均明显增加（P<0.05），ULBP1表达降低（P<0.05）；顺铂诱导A549细胞ATM基因表达明显增加\[(3.23±1.62)×10-6 vs (5.49±3.91)×10-8，P<0.05\]，紫杉醇诱导A549细胞P53基因表达明显增加\[(14.90±5.49)×10-6 vs (3.68±2.82)×10-6，P<0.05\]。结论：顺铂、紫杉醇均可增强CIK细胞对A549细胞的杀伤活性，其分子机制可能与激活DNA损伤修复基因，进而增加NKG2D配体表达有关。

Objective:To study the effect of cisplatin and paclitaxel on the cytotoxic activity of cytokine-induced killer (CIK) cells toward human lung adenocarcinoma cell A549 and further explore the possible molecular mechanism involved. Methods: The IC50 of cisplatin and paclitaxel in inhibiting A549 cells were measured by using MTT assay. Cytotoxicity of CIK cells toward A549 cells that were pre-cultured with cisplatin or paclitaxel was measured through LDH releasing assay. The expression of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) on A549 cells were analyzed by flow cytometry. The level of DNA damage response genes (ATM, ATR, CHK1, CHK2, P53) were assessed by fluorescent quantitative PCR.Results: The IC50 of cisplatin and paclitaxel in inhibiting A594 were 70 μg/ml and 39 μg/ml respectively. Cytotoxicity of CIK cells against A549 cells was significantly increased after the cells were pre-treatment with cisplatin (70 μg/ml) or paclitaxel (39 μg/ml) (P<0.05). Expression of MICA, MICB, ULBP2, and ULBP3 on A549 cells was significantly increased by the cisplatin or paclitaxel pretreatment (P<0.05), where as the level of ULBP1 was decreased after the expose (P<0.05). Furthermore, ATM expression in A594 cells was decreasedby cisplatin treatment \[(3.23±1.62)×10-6 vs (5.49±3.91)×10-8，P<0.05\], whereas the level of P53 mRNA elevated significantly after treatmentwith paclitaxel \[(14.90±5.49)×10-6vs (3.68±2.82)×10-6，P<0.05\]. Conclusion: The results indicated that cisplatinand or paclitaxel can enhance the cytotoxic activity of CIK cells toward lung cancer cells. It is conceivable that they activate DNA-damage response genes, which in turnupregulate the expression of NKG2D ligands that make cells more susceptible to CIK cells.

郑州市科技领军人才项目（No.121PLJRC532）