siRNA沉默HIF—1α在缺氧状态下对胰腺癌细胞TLR4表达的影响  

Effcet of silencing HIF-1α by siRNA on expression of toll-like receptor 4 in pancreatic cancer cells under hypoxic conditions

作　　者：何志强[1] 范平[1] 王博[1] 吴汉青[1] 张景辉[1] 吴河水[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院胰腺外科中心,武汉430022

出　　处：《中华普通外科杂志》2012年第6期475-478,共4页Chinese Journal of General Surgery

基　　金：基金项目：国家自然科学基金资助项目（C160402）

摘　　要：目的观察体外乏氧培养条件下胰腺癌细胞系Panc－1中HIF－1α和TLR4的表达，探讨HIF－1α在低氧条件下对胰腺癌细胞TLR4的调控作用。方法低氧箱培养和CoCl2化学缺氧法模拟肿瘤缺氧环境，RT—PCR、免疫荧光法和流式细胞法分别检测缺氧状态下HIF-1α和TLR4在mRNA和蛋白水平的表达。采用化学合成小干扰RNA（siRNA）介导的RNA干扰技术（RNAi）用siRNA转染Panc－1细胞。观察转染后HIF-1α沉默效果。结果低氧条件下，Panc－1细胞HIF－1α mRNA水平稳定，蛋白表达显著升高，而TLR4mRNA和蛋白的表达均显著升高。siRNA转染Panc－1后能够显著下调HIF－1α的基因表达，同时TLR4基因的表达上调也受到明显抑制。结论缺氧促使胰腺癌Panc－1细胞TLR4在蛋白水平表达升高，TLR4信号通路可能和HIF-1α信号通路存在相互联系，并且共同促进了胰腺癌的恶性进展。Objective To investigate the expression of HIF-1 α and toll-like receptor 4 （TLR4） in human pancreatic cancer cell line pane-1 under hypoxia. To observe the effect of HIF-1α in the regulation of TLR4 expression in pancreatic cancer ceils under hypoxic conditions. Methods Ceils were placed in airtight chamber or treated with CoCl2 to mimic tumor hypoxic micro environment, mRNA and protein levels of HIF-1α and TLR4 were detected by reverse transcription-polymerase chain reaction（RT-PCR） and flow cytometric analysis. By RNA interference （RNAi） originated from small interference RNA （siRNA） to use siRNA was transfeeted into pane-1 cells. Western blot was used to detect gene scilencing effect on HIF-1α. RT-PCR and flow cytometrie analysis was used to observe the change of TLR4 gene expression after HIF-1α gene silence. Results Under hypoxia, both mRNA and protein levels of TLR4 were up-regulated. The siRNA targeting HIF-1 α gene down-regulated HIF-1 α gene in panc-1 cells efficiently, and TLR4 gene was down-regulated as well. Conclusions Hypoxia can increase protein level of TLR4 in pancreatic cancer cells. TLR4 signaling pathway together with HIF-1 α may promote development of the pancreatic cancer.

关 键 词：胰腺肿瘤 Toll样受体4 缺氧诱导因子-1α 亚基 

分 类 号：R735.9[医药卫生—肿瘤]