摘要
目的:观察雷帕霉素(Rapamycin)对人食管癌细胞TE1、TE13中HIF-1α的抑制作用及糖酵解途径相关基因表达的影响;分析mTOR信号通路与糖酵解途径的关系。方法:分别取TE1、TE13细胞株,常氧及缺氧条件下分别予以Rapamycin作用后,将两株细胞各分为四组:①正常对照组,即未处理组(N);②缺氧处理组(H);③常氧Rapamycin处理组(N+R);④缺氧Rapamycin处理组(H+R)(即雷帕霉素预处理1小时后缺氧培养)。采用实时定量PCR检测细胞中缺氧诱导因子-1α(Hypoxia-inducible factor-1α,HIF-1α)蛋白及己糖激酶Ⅱ(HK-Ⅱ)、葡萄糖载体蛋白1(GLUT-1)、乳酸脱氢酶A(LDHA)等糖酵解相关基因mRNA的表达,Western印迹检测HIF-1α及HK-Ⅱ、GLUT-1、LDHA等糖酵解相关基因蛋白的表达。结果:缺氧处理后与正常对照组相比,HIF-1α、HK-Ⅱ、GLUT-1蛋白表达及mRNA明显增强,而LDHA表达无明显变化;Rapamycin预处理后,正常对照组及缺氧处理组HIF-1α、HK-Ⅱ、GLUT-1的表达均明显降低,LDHA无明显变化。结论:常氧及缺氧条件下,雷帕霉素可抑制食管癌细胞HIF-1α及糖酵解相关基因的表达,提示阻断mTOR通路可抑制食管肿瘤细胞的糖酵解途径。
Objective： To investigate the inhibitory effect of Rapamycin on HIF-lct and the impact on glycolysis associated genes in esophageal squamous carcinoma cells. Methods： TEl and TEl3 cell lines were treated with Rapamycin in normoxia or hypox- ia and were divided into four groups respectively： normal control group which was called untreated group （N） ; hypoxia group（H） ; Ra- pamycin treated in normoxia （N ＋ R） ; Rapamycin treated in hypoxia （H ＋ R）. The expression of HIF- 1 oL and glycolysis associated genes （ GLUT- 1, LDHA, HK- 11 ） were measured on mRNA level by reahime RT-PCR and on protein level by Western blot. Results： The expressions of HIF-1 c~, HK- 11 and GLUT-1 mRNA and proteins of TEl and TEl3 cells were upregulated under hypoxia environ- ment compared with that of the normal control group, while no significant change was observed in the expression of LDHA. The expres- sions of HIF- 1 ct, HK- ]] and GLUT- 1 mRNA and proteins in cells treated with Rapamycin were significantly decreased either in nor- moxia or in hypoxia environment. Conclusion： HIF-1 ct may regulate glycolysis through mTOR signaling pathway in esophageal carci- noma ceils.
出处
《肿瘤预防与治疗》
2013年第4期187-191,200,共6页
Journal of Cancer Control And Treatment
基金
国家自然科学基金青年科学基金资助(30800511)
