CIK细胞联合埃克替尼对肺腺癌细胞系的体外抑制效应

In vivo cytotoxicity effects of CIK cells combined with icotinib against lung adenocarcinoma cells

目的观察细胞因子诱导的杀伤细胞(CIK)疗法和盐酸埃克替尼(Icotinib)对肺腺癌PC9细胞和A549细胞的体外杀伤效应。方法以PC9和A549分别为靶细胞,将实验分为4组:CIK细胞单独组、Icotinib单药组、CIK细胞+Ic-otinib联合组和空白对照组。用噻唑蓝(MTT)法检测Icotin-ib对两种靶细胞的半数抑制浓度(IC50);细胞计数(CCK8)试剂盒以及细胞凋亡检测(Annexin V-FITC)试剂盒联合流式细胞仪检测CIK细胞和Icotinib分别及联合对PC9细胞和A549细胞的生长抑制和促凋亡作用。结果①Icotinib对PC9及A549细胞作用48 h后的IC50值分别为:(0.21±0.03)、(4.79±0.74)μmol/L;②CCK8杀伤和Annexin V-FITC凋亡实验可观察:CIK细胞与Icotinib联合应用对靶细胞的体外杀伤作用均明显高于CIK细胞单独组和Icotinib单药组。靶细胞为PC9细胞时,CIK细胞+Icotinib联合组的杀伤率可达(65.85±1.44)%;靶细胞为A549细胞时,CIK细胞+Icotinib联合组的杀伤率可达(52.92±1.57)%。结论 CIK细胞疗法联合分子靶向药物在体外可有效抑制肺癌细胞的生长,分子靶向治疗和细胞免疫疗法为临床上肺癌的综合治疗提供可行性。

Objective To investigate the in vivo effects of cytokine-induced killer （CIK） cells combined with ico- tinib hydrochloride against lung adenocarcinoma cells line PC9 and A549 cells. Methods With PC9 and A549 cells respectively as targeted cells, four groups were divided as follows：CIK cells alone group, icotinib alone group, CIK ＋ icotinib group and blank control group. MTT assay was used to detect the icotinib on two kinds of targeted cells with the half maximal inhibitory concentration （ICs0）. The cytotoxicity and apoptosis of CIK cells combined with icotinib against PC9 cells and A549 cells were respectively determined by Cell Counting Kit（ CCK8 kit）and Annexin V-FITC Apoptosis Detection Kit（ Annexin V-FITC） with flow cytometry. Results （1 At 48 h the IC50 of icotinib on PC9 cells was （0. 21 ±0. 03）μmol/L, while the IC50 of icotinib on A549 cells was （4. 79 ±0. 74） μmol/L; （2） CCK8 damage and Annexin V-FITC apoptosis could be observed that： bination on the effect of killing CIK cells with icotinib corn- targeted cells in vitro were significantly higher than those in the CIK cells alone group and icotinib alone group. From combined group of CIK ＋ icotinib to PC9 cells, the killing rate was （65.85 ± 1.44 ） % , while （52.92± 1.57） % to A549 cells. Conclusion CIK cells combined with icotinib can obviously in- hibit the growth of lung adenocarcinoma cells in vivo. Molecular targeting therapy combined with immunotherapy of- fers the possibility of combined modality therapy in the clinical.