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首页> 中文期刊> 《中国全科医学》 >下调DNA结合分化抑制蛋白1基因对人结肠癌SW480细胞株体外转移和侵袭能力的影响研究

下调DNA结合分化抑制蛋白1基因对人结肠癌SW480细胞株体外转移和侵袭能力的影响研究

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摘要

背景 既往研究已经证实DNA结合分化抑制蛋白1(Id-1)表达升高在结直肠癌的发生、发展中发挥重要作用,但Id-1对结肠癌细胞侵袭行为影响及机制的研究较少.目的 探讨下调Id-1基因对人结肠癌SW480细胞株增殖、转移、侵袭能力的影响及作用机制.方法2016年1-6月,人结肠癌SW480细胞株分为3组:空白组,未进行转染;空载体组,用空载体进行转染;抑制组,采用Id-1特异小干扰RNA (siRNA)进行转染,采用反转录聚合酶链式反应(RT-PCR)和Western blotting法检测各组细胞Id-1和Ki-67基因及蛋白的表达情况,采用MTT法检测细胞增殖能力,采用细胞划痕实验、Transwell小室和Matrigel侵袭实验评价Id-1对细胞转移和侵袭能力的影响.结果 3组细胞Id-1、Ki-67 mRNA及其相应蛋白表达水平比较,差异均有统计学意义(P＜0.05);抑制组细胞Id-1、Ki-67 mRNA及其相应蛋白表达水平较空白组、空载体组降低(P＜0.05).培养第1、2天,3组细胞增殖吸光度(OD值)比较,差异均无统计学意义(P＞0.05).培养第3～7天,3组细胞增殖OD值比较,差异均有统计学意义(P＜0.05);其中抑制组细胞增殖OD值较空白组及空载体组降低(P＜0.05).细胞划痕实验结果显示,抑制组细胞缓慢向中央迁移,缺损处修复较缓慢,空白组和空载体组细胞向划痕处的迁移速度明显加快,而空白组与空载体组间的迁移速度差异不明显.Transwell小室和Matrigel侵袭实验表明,3组迁移细胞数量和侵袭细胞数量比较,差异均有统计学意义(P＜0.05);抑制组迁移细胞数量和侵袭细胞数量较空白组、空载体组减少(P＜0.05).结论 下调Id-1基因能抑制人结肠癌SW480细胞株的增殖、转移和侵袭能力,其机制可能与抑制Ki-67的表达有关.%Background Up-regulation of Id-1 expression playing an important role in the occurrence and development of colorectal cancer has been proved by some previous studies,but down-regulation of it on the invasion of colorectal cancer cells and corresponding mechanism of action are less studied.Objective To explore the effect of deregulated Id-1 expression on the proliferation,metastasis and invasion of human colon carcinoma SW480 cells and investigate its mechanisms of action.Methods This study was conducted from January to June in 2016.SW480 cells with Idl-specific siRNA transfection,no transfection,and blank load transfection were selected as the inhibition group,blank group and blank load transfection group,respectively.Expressions of Id-1 mRNA,Ki-67 mRNA,Id-1 and Ki-67 in these 3 groups were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting.Cell proliferation in vitro was assessed by MTT assay.Effect of Id-1 on SW480 cells was observed by wound healing assay,Transwell migration assay,and Matrigel invasion assay.Results There were significant differences in the expressions of Id-1 mRNA,Ki-67 mRNA,Id-1 and Ki-67 among three groups (P ＜0.05).The expressions of Id-1 mRNA,Ki-67 mRNA,Id-1 and Ki-67 in the inhibition group were lower than those in the blank group and blank load transfection group (P ＜ 0.05).The growth curves of SW480 cells showed that,on the 1st and 2nd days of culture,the differences in the OD value indicating the SW480 cells proliferation ability among three groups were not significant (P ＞0.05),but on the 3rd to the 7th days of culture,the differences in the OD value indicating the SW480 cells proliferation ability among three groups were significant (P ＜ 0.05),and OD value indicating the SW480 cells proliferation ability in the inhibition group was significantly weaker than that in the blank group and the blank load transfection group (P ＜ 0.05).The results of wound healing assay showed that SW480 cells in the inhibition group migrated to the scratched area and closed the wound slowly,while those in the blank group and blank load transfection group did so more quickly,but the difference between the latter two groups was not distinct.Results of Transwell migration assay and Matrigel invasion assay demonstrated that the number of migrating and invading SW480 cells differed significantly among three groups (P ＜0.05);the inhibition group had less the number of migrating and invading SW480 cells than the other two groups (P ＜O.05).Conclusion Deregulated Id-1 has significant inhibition effect on the proliferation,metastasis and invasion of human colon carcinoma SW480 cells.The mechanism of action might be related with the reduced effect of Ki-67.

著录项

  • 来源
    《中国全科医学》 |2017年第8期|953-958|共6页
  • 作者

    武雪亮; 王立坤; 薛军; 杨东东; 屈明; 郭飞; 杨瑞敏; 刘博;

  • 作者单位

    075000河北省张家口市,河北北方学院附属第一医院血管腺体外科;

    075000河北省张家口市,河北北方学院附属第一医院超声科;

    075000河北省张家口市,河北北方学院附属第一医院血管腺体外科;

    075000河北省张家口市,河北北方学院附属第一医院胃肠外科;

    075000河北省张家口市,河北北方学院附属第一医院血管腺体外科;

    075000河北省张家口市,河北北方学院附属第一医院血管腺体外科;

    075000河北省张家口市,河北北方学院附属第一医院超声科;

    075000河北省张家口市,河北北方学院附属第一医院病理科;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 R735.35;
  • 关键词

    结肠肿瘤; 分化抑制蛋白1; 肿瘤转移; 肿瘤浸润;

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