摘要
目的目前细胞因子诱导的杀伤细胞(cytokine-induced kill cell,CIK)逆转肿瘤多药耐药的机制尚不明确,文中旨在研究将CIK细胞与人肺腺癌A549及耐药株A549/DDP细胞非接触式共培养条件下,上清液中细胞因子TNF-α、IFN-γ、IL-2及IL-12的变化情况,为研究CIK细胞逆转A549/DDP耐药机制提供细胞因子水平的理论依据。方法实验将细胞分为5组:CIK组(CIK细胞)、A549组(A549细胞)、A549/DDP组(A549/DDP细胞)、CIK+A549组(CIK细胞与A549细胞按20∶1共培养24 h)、CIK+A549/DDP组(CIK细胞与A549/DDP细胞按20∶1共培养24 h)。利用Transwell小室模型将CIK细胞与A549、A549/DDP细胞分别非接触式共培养后,通过ELISA检测各组细胞培养24 h后上清液中细胞因子TNF-α、IFN-γ、IL-2及IL-12的含量。为测A549/DDP细胞本身耐药倍数,实验将细胞分组:A549a组和A549/DDPa组分别加入A549、A549/DDP细胞,同时加入浓度分别为1、2、4、6、8、16、32、64μg/m L的顺铂;A549 b组、A549/DDP b组分别加入A549细胞、A549/DDP细胞,同时加等体积含5%FBS的RPMI-1640培养液;调零组只加RPMI-1640培养液。为测Transwell中CIK与A549/DDP共培养后A549/DDP耐药倍数,实验将细胞分为对照组(A549/DDP细胞)、实验组(A549/DDP细胞+浓度分别为1、2、4、6、8、16、32、64μg/m L的顺铂)、调零a组(只加培养液)。结果 CIK+A549/DDP组TNF-α含量[(245.82±8.63)pg/m L]显著高于CIK组、A549组、A549/DDP组及CIK+A549组[(52.66±3.32)、(66.95±1.71)、(72.42±3.00)、(88.24±3.44)pg/m L],差异均有统计学意义(P<0.05)。CIK+A549组TNF-α含量较CIK组、A549组、A549/DDP组均升高(P<0.05)。CIK组、CIK+A549/DDP组、CIK+A549组IFN-γ含量显著高于A549组、A549/DDP组(P=0.000)。CIK+A549/DDP组、CIK+A549组IL-2含量[(910.24±32.33)、(502.74±36.24)pg/m L]显著高于CIK组、A549组、A549/DDP组[(144.00±12.14)、(47.32±7.50)、(68.386±2.21)pg/m L],且CIK+A549/DDP组IL-2含量显著高于CIK+A549组,差异有统计学意义(P<0
Objective To study the cytokine-induced killer reversed tumer multi-drug resistance mechanism, to study the non-contact trained CIK cell and chemotherapy sensitive cell line A549 and multidrug resistance cell line A549/DDP, the changes of TNF-ot, IFN-γ, IL-2 and IL-12 in the supernates , secreted by tumor cells. And to explore the evidence for CIK cells reversed A549 / DDP resistance mechanism. Methods CIK cells co-cultured with A549cells , A549/DDP cells respectively in the Transwell model, divided into CIK group, A549 group, A549/DDP group, CIK ＋ A549 group and CIK ＋ A549/DDP group. CIK co-culture with tumor cells according to 20 ： 1 for 24hour. The quantities of cytokines were determined by ELISA. TNF-α, IFN-γ, IL-2 and IL-12 were found in the supernates and analyze them by statistics. A549/DDP cells co- cultured with CIK cells in the Transwell model and that of A549/DDP cells were examined by MTT assay. Results 1. TNF-α, IFN-γ, IL-2 and IL-12 were all found in the supernates of the five groups. 2. The quantities of cytokines were lowest in A549 and A549/ DDP groups, the difference between the two groups was not statistically significant （ P 〉 0.05 ）. 3. After CIK cells non-contact with A549 and A549 / DDP , the quantities of IFN-γ and IL-2 were significantly higher than before, showing statistically significant differences （P 〈 0.05）. 4. After non-contact co-culture with CIK cells, the quantities of TNF-α, IL-2 and IL-12 in CIK ＋ A549/DDP were significantly higher than those in CIK ＋ A549 group, showing statistically significant differences （P 〈 0.05 ）. 5. MTT assay shows the resistent factor of A549/DDP cells （ RF1 ） = 5. 087, that of A549/DDP cells co-cultured in Transwell model （ RF2 ） = 0. 805. Conclusion By CIK non-contact co-cultured with tumor cells, especially the drug-resistant strain A549/DDP co-cuhured with CIK, the secretion of TNF-α,IL-2 and IL-12 were more significantly stimulated. The reversal fold of A549/DDP cells decreases after the co- culture of
出处
《医学研究生学报》
CAS
北大核心
2016年第1期29-33,共5页
Journal of Medical Postgraduates
基金
国家自然科学基金(81260397)
贵州省科技厅科学技术基金[黔科合J字(2013)2331号]
贵州省高校优秀科技创新人才支持计划基金[黔教合KY字(2015)495]
遵义医学院博士启动基金(2012-2016)
遵义市科技局省-市联合项目基金[遵科合人(2014)9号]
