siRNA干扰Id-1表达对人结肠癌SW480细胞株生长与侵袭能力影响研究  

Impact of siRNA interference reducing Id-1 on invasion and metastasis of human colon carcinoma cell line SW480 in vitro

作　　者：武雪亮[1] 王立坤[2] 杨东东[1] 薛军[1] 屈明[1] 郭飞[1] 刘博[3] 

机构地区：[1]河北北方学院附属第一医院普外科,河北张家口075000 [2]河北北方学院附属第一医院超声医学科,河北张家口075000 [3]河北北方学院附属第一医院病理科,河北张家口075000

出　　处：《中华肿瘤防治杂志》2017年第3期156-160,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：河北省卫计委医学科学研究重点课题计划(20150058);河北省科技支撑计划(152777237)

摘　　要：目的 DNA结合分化抑制因子-1(inhibitor-1of DNA binding/differentiation-1,Id-1)对结肠肿瘤细胞侵袭行为影响的研究较少,本研究探讨siRNA转染下调Id-1表达对人结肠癌SW480细胞侵袭和转移行为的影响。方法将合成Id-1干扰序列(siRNA)转染至SW480细胞作为实验组(抑制组),并以Id-1表达干扰空白组、空载体组作为对照,应用反转录聚合酶链反应(RT-PCR)和蛋白质印迹法检测各组细胞中Id-1和基质金属蛋白酶-9(matrix metalloprotein,MMP-9)的表达情况,四甲基偶氮唑盐(MTT)法检测细胞的增殖能力;划痕损伤实验、Transwell小室和Matrigel侵袭模型评价Id-1对细胞侵袭和转移能力的影响。结果抑制组Id-1mRNA表达水平为(0.14±0.02),与空白组(1.27±0.03)和空载体组(1.25±0.06)比较明显降低,差异有统计学意义,P<0.001;抑制组Id-1蛋白表达水平为0.25±0.02,与空白组(1.18±0.03)和空载体组(1.16±0.04)比较明显降低,差异有统计学意义,P<0.001;抑制组MMP-9mRNA表达水平为0.19±0.02,与空白组(1.33±0.04)和空载体组(1.38±0.03)比较明显降低,差异有统计学意义,P<0.001;抑制组MMP-9蛋白表达水平为0.12±0.02,与空白组(0.89±0.04)和空载体组(0.91±0.02)比较明显降低,差异有统计学意义,P<0.001;生长曲线表明,从第3天起抑制组细胞增殖明显减弱,与空白组和空载体组差异明显;划痕损伤实验结果显示,空白组和空载体组向划痕处的迁移速度明显快于抑制组;迁移试验显示,抑制组迁移细胞数为75±12,明显少于空白组(201±12)和空载体组(206±15);抑制组侵袭细胞数为51±10,明显少于空白组(121±17)和空载体组(126±14),差异均有统计学意义,P<0.001。结论 Id-1促进SW480细胞的侵袭和转移,其机制可能与诱导MMP-9的表达有关。OBJECTIVE There is less research with the invasion and metastasis of Inhibitor-1of DNA binding/differentiation-1（Id-1）in colorectal cancer.This study investigates the influence on invasion and metastasis of human colon carcinoma cell line SW480 in vitro by siRNA interference reducing Id-1.METHODS SW480 cells transfected with Id-1interference sequence（siRNA）were selected as the experimental group（the inhibition group）,and SW480 cells with blank interference sequence and blank load transfection group were selected as the control group.Expression of Id-1and Matrix metalloprotein（MMP-9）in three groups were detected by reverse transcription polymerase chain reaction（RT-PCR）and Western blot.Cell proliferation was assessed by methyl thiazolyl tetrazolium（MTT）assay.Cell invasion and migration of Id-1was observed by Wound healing,Transwell chamber experiment and Matrigel invasion assay.RESULTS The expression of Id-1 mRNA in the inhibition group（0.14±0.02）was lower than those in the blank group（1.27±0.03）and the blank load transfection group（1.25±0.06）,and there was significant difference（P〈0.001）.The results of Western blot demonstrated that the expressions of Id-1protein in the inhibition group（0.25±0.02））was lower than those in the blank group（1.18±0.03）and the blank load transfection group（1.16±0.04）,and there was a significant difference（P=0.000）.The expressions of MMP-9mRNA in the inhibition group（0.19±0.02）was lower than those in the blank group（1.33±0.04）and the blank load transfection group（1.38±0.03）,and there was a significant difference（P=0.000）.The expressions of MMP-9protein in the inhibition group（0.12±0.02）was lower than those in the blank group（0.89±0.04）and the blank load transfection group（0.91±0.02）,and there was a significant difference（P=0.000）.The growth curve showed that proliferation ability in the inhibition group was significantly weaker than that in the blank group and the blank load transfect

关 键 词：结肠肿瘤 DNA结合分化抑制蛋白1 侵袭 转移 

分 类 号：R735.35[医药卫生—肿瘤]