CIK细胞联合多西紫杉醇对肺腺癌耐药细胞SPC-A1/DTX的体内外杀伤作用的实验研究  

The anti-tumor effects of CIK cells combined with docetaxel against drug-resistant lung adenocarcinoma cell line SPC-A1/DTX in vitro and in vivo

作　　者：刘鹏英[1] 陈龙邦[1] 

机构地区：[1]南京大学医学院临床学院(南京军区南京总医院)肿瘤内科,南京210002

出　　处：《中国免疫学杂志》2008年第12期1079-1083,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.30872929);南京军区南京总医院重点科研基金项目(Z2008025)

摘　　要：目的:探讨正常人细胞因子诱导的杀伤(Cytokine induced-killer,CIK)细胞联合多西紫杉醇对肺腺癌多西紫杉醇耐药细胞SPC-A1/DTX的体内外杀伤作用。方法:取健康人外周血单个核细胞,经IFN-γ、IL-2、抗CD3单抗联合诱导CIK细胞,用四甲基偶氮唑蓝(MTT)法测定多西紫杉醇、CIK细胞及两者联合对耐药细胞SPC-A1/DTX体外的细胞毒活性。用肺腺癌耐药细胞SPC-A1/DTX建立裸鼠皮下移植瘤模型,2周后腹腔给予生理盐水(对照组)、多西紫杉醇(单纯化疗组)、CIK细胞和CIK细胞联合多西紫杉醇(联合治疗组),观察它们对裸鼠移植瘤生长的抑制作用。结果:体外实验表明,SPC-A1/DTX对多西紫杉醇的耐受性(IC50为110.5μg/ml)较亲代敏感株SPC-A1(IC50为8.5μg/ml)增加了13倍。CIK细胞对人肺腺癌多西紫杉醇耐药细胞SPC-A1/DTX的杀伤活性,明显高于其亲代敏感株SPC-A1(P<0.05)。经CIK细胞作用后的SPC-A1/DTX细胞,仅10μg/ml的DTX即可使CIK:SPC-A1/DTX效靶比为5:1组中的联合杀伤率比单纯加DTX(同等剂量)组增加6.1倍,比单纯加CIK(相同效靶比)细胞杀伤率增加1.4倍,联合杀伤率的大小随效靶细胞比值和DTX浓度的升高呈依赖性增加。体内实验表明,CIK细胞联合多西紫杉醇可明显抑制裸鼠皮下肺腺癌耐药细胞移植瘤的生长,联合治疗组肿瘤生长缓慢,肿瘤组织坏死、淋巴细胞浸润明显。结论:CIK细胞联合多西紫杉醇能够显著抑制体内外肺腺癌多药耐药细胞SPC-A1/DTX的生长,为临床上应用CIK细胞联合化疗治疗放、化疗等无效的中晚期耐药肺癌患者提供了实验依据。Objective：To investigate the inhibitory effects of cytokine induced-killer（CIK） cells combined with docetaxel（DTX） on the expansion of lung adenocarcinoma cell line SPC-A1/DTX in vitro and in vivo.Methods：Peripheral blood mononuclear cells（PBMC） from healthy donors were incubated in vitro to induce CIK cells in the presence of interferon-gamma（IFN-γ）,IL-2 and anti-CD3 monoclonal antibody.MTT assay was employed to evaluate the cytotoxic activity of DTX,CIK cells and their combination against SPC-A1/DTX cells in vitro.SPC-A1/DTX lung adenocarcinoma cells were injected subcutaneously into nude mice.On the 14 th day,normal saline（control group）,docetaxel（DTX 1 mg/kg in 0.2 ml,DTX group）,CIK cells（1×10^7,CIK group）,and CIK cells combined with docetaxel（CIK＋DTX group） were administered intraperitoneally respectively.All the nude mice were sacrificed at day 15 after treatment and the tumor were weighted out.Results：MTT assay showed that the IC50 to docetaxel in SPC-A1/DTX cells was 110.5 μg/ml and 8.5 μg/ml in wild SPC-A1 cells.CIK cells possessed a higher anti-tumor cytotoxic activity in SPC-A1/DTX cells in vitro than in wild SPC-A1 cells（P〈0.05）.The killing activity of CIK cells combined with DTX was 1.4 times more than that of separate use of CIK cells,and 6.1 times more than that of separate use of DTX.Increasing the E∶T ratio and the concentration of docetaxel directly correlated with higher mean percent specific lysis.In addition,CIK cells plus DTX had a stronger suppressive effect on the tumor growth in nude mice bearing SPC-A1/DTX tumor in vivo.Compared to other groups,in the combined therapy group of mice,the tumor grew slowly,with more marked tissue necrosis and more infiltrated lymphocyte.Conclusion：CIK cells combined with docetaxel can significantly inhibit the growth of MDR cells of lung cancer in vitro and in vivo.The result provides an experimental basis for CIK combined with chemotherapy for clinical application in treatment of MDR lung cancer.

关 键 词：肺癌 多药耐药 过继性细胞免疫治疗 细胞因子诱导的杀伤细胞 肺腺癌耐药细胞系SPC-A1/DTX 

分 类 号：R730.51[医药卫生—肿瘤]