摘要
目的研究基因沉默缺氧诱导因子-1α(HIF-1α)和不同缺氧条件下HIF-1α、锌指转录因子(Snail)及上皮钙黏蛋白(E-cadherin)在SW480细胞中的表达,及其对生长增殖和侵袭迁移力的影响。方法构建SW480细胞缺氧模型,设计合成HIF-1α的小干扰RNA并将其转染入细胞内于无氧环境培养。采用MTT法检测生长增殖活性,Transwell法检测体外侵袭迁移能力,RT-PCR检测HIF-1α、Snail及E-cadherin mRNA的表达。结果 MTT显示无氧组和阴性对照组生长增殖活性增加,基因干扰组降低(P<0.05);体外侵袭实验显示无氧组、阴性对照组侵袭迁移率增加,基因干扰组降低(P<0.05);RT-PCR检测显示无氧组、阴性对照组HIF-1α和Snail mRNA表达升高,E-cadherin降低,基因干扰组HIF-1α和Snail表达降低,E-cadherin升高(P<0.05),且相关分析显示HIF-1α与Snail呈正相关,HIF-1α与E-cadherin呈负相关。结论缺氧及HIF-1α可促进SW480细胞株的体外生长增殖活性,增强其体外侵袭迁移能力,基因沉默HIF-1α可抑制其体外生长增殖与侵袭迁移。
[Objectives] To study gene silencing hypoxia inducible factor-1α （HIF-1α） and the expres- sion level of HIF-1α, Snail and E-cadherin in different hypoxia condition in the SW480 ceils, and research its effect of the proliferation, invasion and transference. [Methods] Chemical induced the cell hypoxia model, we designed and synthetized the HIF-lct-siRNA, and got the cells transfeetion, the groups was divided into： normoxia, hypoxia, negative control （hypoxia） and gene interference group （hypoxia）. MTT was used to detect the proliferation of the SW480 cells, transwell was applied to evaluate the invasion and transference capability of the ceils, the mRNA expression levels of HIF-I ct, Snail and E-cadherin were detected by RT-PCR. [Results] MTT disclosed that the proliferation of the cells were increased in hypoxia and negative control groups, the gene interference group was decreased （P〈0.05）; Transwell showed that the migration rate was in- creased in hypoxia and negative control groups, the gene interference group was decreased （P〈0.05）; RT-PCR displayed that the cells＂ mRNA and protein expression of HIF-lc~ and Snail were increased and E-cadherin were decreased in hypoxia and negative control groups, the cells＂ mRNA and protein expression of HIF-lct and Snail were decreased and E-cadherin were increased in gene interference group （P 〈0.05）, furthermore, HIF-1α had positive correlation with Snail, and negative correlation with E-cadherin （P〈0.05）. [Conclusion]The hypoxia and HIF-1α can promote the extraorgan proliferation activities of the SW480 cells, enhance its invasion and migration activities, and gene silencing HIF-1α can inhibit it.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2013年第29期41-45,共5页
China Journal of Modern Medicine
基金
衡阳市科技局资助项目(No:2010kj50)
