代谢基因ARG1对结直肠癌细胞增殖和凋亡的影响  

Effect of metabolic gene ARG1 on cell proliferation and apoptosis of human colorectal cancer

作　　者：黄娟妮[1] 林绪涛[2] 范德军[2] 梁丽英[1] 陈广原[1] 叶慧玲[1] 

机构地区：[1]广州医科大学附属第一医院老年病科,广州510120 [2]中山大学附属第六医院结直肠肛门外科

出　　处：《中华生物医学工程杂志》2015年第2期119-124,共6页Chinese Journal of Biomedical Engineering

基　　金：广州医科大学青年科研项目（2013A05）

摘　　要：目的：探讨精氨酸酶1（ARG1）对人结直肠癌细胞增殖和凋亡的影响。方法应用小干扰RNA转染HT29细胞沉默ARG1表达，应用慢病毒载体转染HT29细胞高表达ARG1基因；与巨噬细胞M2细胞共培养；应用real time RT-PCR检测各组细胞ARG1基因表达水平；应用精氨酸酶活性实验检测各组细胞ARG1活性。应用ELISA法检测各组培养液中IL-4及IL-6浓度。CCK8及TUNEL试剂盒检测各组HT29细胞增殖及凋亡情况。结果与对照组相比，外源性添加L-精氨酸或高表达ARG1基因后，HT29细胞ARG1活性、IL-4与IL-6分泌水平及细胞增殖水平均显著提高（P〈0.05），而HT29细胞凋亡水平则显著降低（P〈0.05）；而外源性添加精氨酸酶抑制剂或应用小干扰RNA沉默ARG1基因表达后，HT29细胞ARG1活性、IL-4与IL-6分泌水平及细胞增殖水平均显著降低（P〈0.05），而凋亡水平则显著提高（P〈0.05）。结论 ARG1可通过促进肿瘤免疫微环境中精氨酸降解，诱导肿瘤相关巨噬细胞因子IL-4及IL-6的分泌，从而促进结直肠癌细胞的增殖并抑制其凋亡。ARG1有可能成为结直肠癌一个新的治疗靶点。Objective To investigate the effect of arginase1 （ARG1） on cell proliferation and apoptosis of human colorectal cancer （CRC）. Methods ARG1gene expression was silenced and over-expressed by transfection of HT29 cells with small interfering RNA （siRNA） and lentiviral vector, respectively. The transfected cells were co-cultured with macrophage M2. The ARG1 gene expression level of the cells in each group was measured by real time RT-PCR. The ARG1 activity of the cells in each group was determined by arginase activity assay. The concentrations of IL-4 and IL-6 in the medium of each group were measured by enzyme-linked immunosorbentassay（ELISA）. Cell proliferation and apoptosis of HT29 in each group were assessed by the CCK-8 and TUNEL assay kits,respectively. Results Compared with the control group,the ARG1 activity,IL-4 and IL-6 secretion levels,and proliferation of HT29 cells significantly increased after adding exogenous L-arginine（L-Arg）or high expression of ARG1 gene（P〈0.05）,whereas the apoptosis of HT29 cells significantly decreased （P〈0.05）. However, after adding exogenousarginaseinhibitor or applying siRNA to silence the ARG1 gene expression,the ARG1 activity,IL-4 and IL-6 secretion levels,and proliferation of HT29 cells significantlydecreased （P〈0.05）,whereas the apoptosis of HT29 cells significantly increased（P〈0.05）. Conclusion ARG1 can induce the secretion of tumor-associated macrophages cytokines IL-4 and IL-6 by promoting arginine degradation in tumor-immune无nbsp;microenvironment,thereby promoting the proliferation and inhibiting the apoptosis of CRC cells. ARG1 may be a new therapeutic target for CRC.

关 键 词：结直肠肿瘤 细胞增殖 细胞凋亡 

分 类 号：R735.34[医药卫生—肿瘤]