摘要
目的探讨炎症微环境下ARG1基因表达水平对人结直肠癌细胞增殖的影响。方法选取人结直肠癌细胞系HT29细胞和巨噬细胞M2型细胞,通过小干扰RNA转染HT29细胞沉默ARG1表达,通过慢病毒载体系统转染HT29细胞高表达ARG1基因。实验共分6组:HT29单纯培养组、HT29与M2细胞共培养组、共培养+精氨酸酶抑制剂组、共培养+L-精氨酸组、ARG1高表达HT29与M2细胞共培养组、ARG1沉默HT29与M2细胞共培养组。精氨酸酶活性实验检测各组细胞中ARG1活性,ELISA法检测各组培养液中IL-6含量,CCK8试剂盒检测各组HT29细胞增殖能力。结果与巨噬细胞M2型细胞共培养后HT29细胞中精氨酸酶活性明显高于单独培养组。外源性添加L-精氨酸或高表达ARG1基因后,HT29细胞精氨酸酶活性、IL-6分泌水平及细胞增殖水平均显著提高。外源性添加精氨酸酶抑制剂(Nor-NOHA)或转染si RNA沉默ARG1基因表达均能明显降低HT29精氨酸酶活性,IL-6分泌水平及细胞增殖水平均显著降低。结论炎症微环境下过表达代谢基因ARG1能够增强结直肠癌细胞HT29的增殖能力。
Objective To investigate the effect of ARG1 gene expression on proliferation of human colorectal cancer cell lines under inflammatory microenvironment. Methods Human colorectal cancer cell line HT29 cells and macrophage M2 cells were chosen for co-culture. si RNA was transfected into to silence ARG1 gene expression and lentivirus vector system was transfected into HT29 cells to induce high expression of ARG1 gene. Experiment groups were divided into six groups, only HT29 group, HT29+M2 cell co-culture group, co-culture +arginine enzyme inhibitor group, co-culture of +L-arginine group, high ARG1 expressed HT29 cell and M2 cell co-culture group, ARG1 silencing HT29 and M2 cell co-culture group. The activity of ARG1 was detected with arginine acid enzyme activity experiment. IL-6 level was detected by ELISA.CCK8 kit was used to detect proliferation of HT29 cells. Results HT29 cell arginase activity was significantly higher when HT29 cells were co-cultured with macrophage M2 cells. Exogenous addition of Larginine or high expression of ARG1 gene significantly increased HT29 cell arginase activity, IL-6 secretion level and cell proliferation.Exogenous addition ofarginine enzyme inhibitor or silencing ARG1 gene expressionsignificantlydecreased HT29 cell arginase activity, IL-6 secretion level and cell proliferation.Conclusion Under inflammatory microenvironment, overexpression of ARG1 genes can enhance the proliferation of colorectal cancer HT29 cells.
出处
《消化肿瘤杂志（电子版）》
2015年第1期33-37,共5页
Journal of Digestive Oncology(Electronic Version)
