摘要
目的 探讨细胞因子从大肠癌癌患者外周血贴壁单个核细胞（PBMCs）诱导的树突状细胞（DC）与杀伤细胞（CIK）对人大肠癌SW620细胞增殖的作用及分子机制.方法 用大肠癌患者大肠癌组织裂解物（肿瘤抗原）致敏经粒细胞-巨噬细胞集落刺激因子（GM-CSF）、肿瘤坏死因子（TNF）-α及白细胞介素（IL）-4等诱导PBMC产生的DC.用干扰素（IFN）-γ、IL-2和人CD3单克隆抗体（hCD3mAb）等诱导该PBMC的悬浮细胞产生CIK细胞.用6孔板将DC-CIK细胞与SW620细胞在同一培养体系内经分别培养24、48、72 h.以噻唑蓝（MTT）法检测癌细胞增殖；以抗酒石酸酸性磷酸酶（TRAP）-酶联免疫吸附试验（ELISA）法检测癌细胞端粒酶活性；采用Western blot法检测人端粒酶逆转录酶（hTERT）蛋白.结果 大肠癌细胞经DC-CIK处理72 h后,空白对照组和DC-CIK-a 、DC-CIK-b组相对增殖率分别为（95.6±2.2）％、（73.5±1.8）％、（51.8±1.6）％；端粒酶活性检测发现,空白对照组24、48、72 h端粒酶活性分别为1.568±0.078、1.535±0.056和1.527±0.039;DC-CIK-a组24、48、72 h端粒酶活性分别为1.356±0.056、1.108±0.039和0.578±0.028；DC-CIK-b组24、48、72 h端粒酶活性分别为1.181 ±0.075、0.718±0.039、0.386±0.036.Westernblot检测发现,与空白对照组比较,DC-CIK-a组和DC-CIK-b组hTERT蛋白水平明显下调.结论 DC-CIK细胞可明显抑制大肠癌细胞增殖；抑制端粒酶活性是其重要机制之一.
Objective To explore the effects and mechanism of cytokine induced killer （CIK）cells and dendritic cells （DC） from peripheral blood mononuclear cells （PBMC） of patients with human colorectal cancer on prloferation and apoptosis of human liver cancer cell line SW620.Methods Adherent PBMC were induced by granulocyte macrophage-colony stimulating factor （GM-CSF）,tumor necrosis factor （TNF）-α and interleukin （IL）-4 to prepare DC.CIK cells were obtained from suspended PBMC induced by interferon （IFN）-γ,IL-2 and human CD3 monoclonal antibody.The colorectal cancer SW620 cells that were cultured without DC-CIK cells served as the controls.DC-CIK cells and SW620 cells were cultured in a well cell cultrue cluster for 24,48 and 72 h,respectively.The proliferation of SW620 cells that were cultured with DC-CIK cells for 24,48 and 72 h was detected by methyl thiazol tetrazolium （MTT） assay.The telomerase activity was determined by tartrate resistant acid phosphatase （TRAP）-enzyme linked immunosorbent assay （ELISA）.Human telomerase reverse transcriptase （hTERT） gene was evaluated by Western blotting.Results In colorectal cancer cells treated with DC-CIK after 72 h treatment,cell relative growth rate in control group,and DC-CIK-a group and DC-CIK-b group was （95.6 ± 2.2） %,（73.5 ±1.8）% and （51.8 ± 1.6）% respectively.The results from TRAP-ELISA showed that telomerase activity in control group at 24,48 and 72 h was 1.568 ±0.078,1.535 ±0.056,and 1.527 ±0.039,that in DCCIK-a group was 1.356 ±0.056,1.108 ±0.039,and 0.578 ±0.028,and that in DC-CIK-b was 1.181 ±0.075,0.718 ± 0.039,and 0.386 ± 0.036,respectively.The Western blotting results showed that the hTERT protein in DC-CIK-a group and DC-CIK-b group was down-regulated as compared with control group.Conclusion DC-CIK cells could inhibit the proliferation of colon cancer cells through down-regulating telomerase activity.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第11期2302-2304,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81250035）
江苏省＂333工程＂科研基金资助项目（BRA2012103）
江苏省创新基金资助项目（CXLX12-0679）
江苏大学临床科技基金资助项目（JLY2012006）
关键词
大肠癌
树突状细胞-杀伤细胞
增殖
端粒酶
Colorectal cancer
Dendritic cell-cytokine induced killer
Proliferation
Telomerase
作者简介
通信作者：范钰，Email：yuf36@sina．com
