摘要
目的探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)中低氧诱导因子-1α(hypoxia-inducible fac-tor-1α,HIF-1α)对血管生成素-2(angiopoietin-2)表达的影响及意义。方法免疫组化SP法检测46例NSCLC组织HIF-1α、血管生成素-2蛋白的表达;培养人肺腺癌细胞株A549细胞,CoCl2模拟缺氧处理6h、12h、24h,以及缺氧条件下不同浓度genistein处理细胞12h后,采用Western Blot和RT-PCR方法分别检测A549细胞HIF-1α蛋白和血管生成素-2 mRNA的表达情况。结果46例NSCLC中HIF-1α、血管生成素-2的阳性表达率分别为73.9%(34/46)、63.0%(29/46),明显高于癌旁肺组织(P<0.05);HIF-1α的表达与血管生成素-2的表达呈正相关。急性缺氧可以诱导A549细胞HIF-1α蛋白和血管生成素-2 mRNA表达增加,分别在6h,12h达到高峰,随着缺氧时间延长,HIF-1α蛋白和血管生成素-2 mRNA表达相对减少;genistein抑制HIF-1α蛋白后,血管生成素-2 mRNA的表达也相应减少,呈浓度依赖。结论缺氧时NSCLC中HIF-1α可参与血管生成素-2的调控,HIF-1α、血管生成素-2表达增加与肿瘤新生血管形成有关,二者共同促进NSCLC的发生发展过程。
Objective To investigate the effect of hypoxia-inducible factor-1 α on angiopoietin-2 expression in NSCLC. Methods The expressions of HIF-1α and angiopoietin-2 were detected in 46 human NSCLC samples by SP immunohistochemistry. The productions of HIF-1α protein and angiopoietin-2 mRNA were observed at different hypoxia culture phases （6h, 12h, and 24h） and in the presence of different doses of genestein. HIF-1α protein was tested by Western Blot analysis, and the expression of angiopoietin-2 mRNA was shown using the reverse transcription PCR （RT-PCR） . Results The positive rates of HIF-1α and angiopoietin-2 were significantly higher in human NSCLC than those in paraneoplastic tissues （P 〈 0. 05 ） ; and HIF-1α expression was positively related to the expression of angiopoietin-2. Both HIF-1α protein and angiopoietin-2 mRNA were hypoxia-induced in a timedependent manner, at 6h and reaching HIF-1α protein expression peaking at 6h, and angiopoietin-2 expression beginning to increased the maximum at 12h of treatment. But genestein treatment could partially counteract the hypoxi- a-induced angiopoietin-2 expression. Conclusion In NSCLC, the HIF-1α protein may regulate giopoietin-2 under hypoxic conditions, and the increased expressions of HIF-1α protein and are correlated with tumor angiogenesis and contribute together to the development of NSCLC. the expression of anangiopoietin-2 mRNA
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2007年第4期451-456,共6页
Chinese Journal of Histochemistry and Cytochemistry
