摘要
目的探讨缺氧环境对人胰腺癌细胞株PANC1诱导因子-1α(hypoxia-inducible factor1α,HIF-1α)、整合素α5(integrina,INFα5)和整合素β1(integrinβ1,INTβ1)表达及侵袭力的影响。方法以5%氧条件下培养PANC1,采用RT-PCR和Western blot检测HIF-1α、INTα5、INTβ1的mRNA和蛋白表达;采用Matrigel胶观测人胰腺癌细胞黏附侵袭能力的变化。并加入HIF-1a抑制剂YC-1缺氧培养PANC1,观察上述各指标的变化。结果HIF-1α蛋白表达从常规培养时0.497±0.011逐渐上升到缺氧培养24h的1.455±0.133(P〈0.05),应用YC-1后蛋白表达下降到0.465±0.073(P〈0.05),而HIF-1α mRNA表达无明显变化;INTα5蛋白从常规培养时0.964±0.032逐渐下降到缺氧培养24h的0.465±0.073(P〈0.05),应用YC-1后蛋白表达又上升到0.883±0.013(P〈0.05),INTα5mRNA的表达从0.212±0.023下降到0.018±0.002(P〈0.05),应用YC-1后再上升到0.069±0.011(P〈0.05);INTβ1的蛋白和mRNA表达于缺氧培养及YC-1处理后均无明显变化。细胞侵袭力从常规培养时69.9±30.5逐渐上升到缺氧培养24h的105.0±14.9(P〈0.05),应用YC-1后又下降到79.7±20.9(P〈0.05)。结论缺氧微环境下HIF-1α过表达可能通过下调INTα5来调控胰腺癌细胞间和细胞与基质间的黏附能力,有利于进一步侵袭转移。
Objective To investigate the effect of hypoxia on expression of HIF-lα and integrinαs β1 invasiveness and in pancreatic carcinoma cell line PANC1. Methods The expression of HIFlα, INTαs, INTβ1 protein in PANC1 cells was examined by Western blot; The expression of their mRNA was delected by RT-PCR. The invasive ability of cells was measured by the number of cells to penetrate polycarbonates coated with matrigel. Results HIF-lα protein expression of PANC1 increased when cells were exposed to hypoxia and reached peak at hypoxia 24 hours (P 〈 0.05). The expression of INTα5 mRNA and protein reduced significantly after hypoxia cuhrse and kepted declining with the time of hypoxia prolonged(P 〈 0.05). YC1 could inhibit the upregulation of HIF-lα, but INTα5 protein in hypoxia environment (P 〈 0. 05) . The expression of INTβ1 mRNA and protein was markedly increased. Invasive tests also showed that the invasive ability of PANC1 cells was inhibited when the expression of HIF-lα inhibited by YC-I (P 〈 0.05). Conclusions HIF-la may be an upstream regulator of INTα5 and HIF-lα over expression, and play an important role in metastasis of pancreatic cancer.
出处
《胰腺病学》
2007年第3期181-183,共3页
Chinese JOurnal of Pancreatology