TIMP1基因对人结肠癌HCT-8细胞增殖影响的研究  

Effect of TIMP1 Gene in Proliferation of Human Colon Carcinoma Cells

作　　者：马艳娇[1,2] 李鹏[1,2] 宫鹏涛[1] 杨举[1] 李赫[1] 李建华[1] 欧阳红生[1] 张西臣[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]黑龙江八一农垦大学动物科技学院

出　　处：《环境与健康杂志》2012年第2期146-149,F0003,共5页Journal of Environment and Health

基　　金：国家重大基础研究计划资助项目(2006CB910505)

摘　　要：目的采用体外试验探讨基质金属蛋白酶抑制剂1(TIMP1)基因对人结肠癌(HCT-8)细胞增殖的影响。方法构建TIMP1基因的真核表达载体和RNA干扰质粒(pcDNA3.1-TIMP1-K、TIMP1 RNAi、pcDNA3.1和空白对照),将TIMP1基因真核表达质粒和RNA干扰质粒分别瞬时转染入HCT-8细胞48～72 h后,采用Real Time PCR方法检测TIMP1基因在HCT-8细胞中基因转录表达水平,并用Western blot方法检测TIMP1基因在HCT-8细胞中蛋白表达水平,采用MTT试验检测TIMP1基因过表达和干扰后对HCT-8细胞增殖的影响。结果成功构建了TIMP1真核表达质粒和RNA干扰质粒。与空白对照比较,质粒pcDNA3.1-TIMP1-K在HCT-8细胞中mRNA和蛋白的相对表达含量较高,质粒TIMP1 RNAi的mRNA和蛋白表达含量较低,差异均有统计学意义(P<0.05)。构建过表达质粒(pcDNA3.1-TIMP1-K)转染HCT-8细胞后,与空白对照相比,第1～4天HCT-8细胞的活性无显著变化(P>0.05),第5～7天TIMP1基因过表达能够促进HCT-8细胞的增殖活性(P<0.05);构建干扰质粒(TIMP1 RNAi)转染HCT-8细胞后,与空白对照相比,第1～4天HCT-8细胞生长无显著变化(P>0.05),第5～7天TIMP1基因过表达能够抑制HCT-8细胞增殖(P<0.05)。结论 TIMP1基因过表达后能够促进人结肠癌细胞的增殖,在一定条件下还能够抑制人结肠癌细胞的增殖。Objective To explore the effects of TIMP1 in proliferation of colon carcinoma cells. Methods The pcDNA3.1-TIMPI-k plasmids and interfere plasmid were constructed and the human colon carcinoma cells （HCT-8） were transfected by them. After 48-72 h,the transcriptional level and protein level were detected by the method of real time PCR and western blot,while the growth of the HCT-8 cells was also validated by MTF. Results The pcDNA3.1-TIMPI-k plasmids and interfere plasmid were constructed successfully, while the TIMP1 gene expression was enhanced by TIMP1 over-expression plasmid and degraded by TIMP1 interfere RNA plasmid. For the four days ,there was no significant difference（P〉0.05 ） in MTr. At the fifth day ,there was significant difference （P〈0.05）, yet at the sixth and seventh days ,there was significant difference （P〈0.01）. The results showed that the transcriptional and protein level were reduced by the TIMP1 interfere RNA plasmid,yet the transcriptional and protein level were elevated by the TIMP1 over-expression plasmid. While it was demonstrated that the low-expression of TIMPI could inhibit the growth of the HCT-8 cells and the over-expression of TIMPI could improve the growth of cells from the growth curve picture. Conclusion Up-regulated expression of TIMP1 can promote the proliferation of HCT-8 cells.

关 键 词：基质金属蛋白酶抑制剂1 人结肠癌细胞 增殖 

分 类 号：R994.6[医药卫生—毒理学]