顺铂和紫杉醇对CIK细胞杀伤肺癌A549细胞的影响及其可能的机制  

The effect of cisplatin or paclitaxel on the activity of CIK cells to kill lung cancer A549 cells and its possible mechanism

作　　者：徐虹[1] 梅家转[1] 

机构地区：[1]南方医科大学附属郑州人民医院肿瘤内科,河南郑州450003

出　　处：《中国肿瘤生物治疗杂志》2015年第4期443-447,共5页Chinese Journal of Cancer Biotherapy

基　　金：郑州市科技领军人才项目(No.121PLJRC532)~~

摘　　要：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(IC50),LDH释放法检测IC50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IC50分别为70、39μg/ml。IC50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)×10-6vs(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)×10-6vs(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 IC50 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 NKG2 D 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 IC50 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- 6vs( 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- 6vs( 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 NKG2 D ligands that make cells more susceptible to CIK cells.

关 键 词：非小细胞肺癌 顺铂 紫杉醇 CIK细胞 NKG2D配体 DNA损伤修复基因 

分 类 号：R734.2[医药卫生—肿瘤]