摘要
目的:观察转染缺氧诱导因子1α(HIF-1α)对人前列腺癌细胞血管形成相关蛋白的影响,并探讨其分子机制。方法:用脂质体Lipofectamine2000包装重组真核表达载体pCDNA3.1(-)/HIF-1α后转染人前列腺癌细胞LNCaP,600μg/ml G418筛选稳定表达的抗性克隆。免疫荧光及Western印迹法鉴定HIF-1α过表达,Western印迹法检测血管形成相关蛋白血管内皮生长因子(VEGF)、iNOS、促血管生成素2(Ang-2)。结果:与未转染细胞LN-CaP相比,转染细胞LNCaP/HIF-1α中出现明显的HIF-1α蛋白条带,并激发出较强荧光,VEGF、iNOS表达增加,Ang-2未见明显变化。结论:HIF-1α过表达能够诱导LNCaP细胞血管形成相关蛋白表达增多,进而增强其体外血管形成能力。
Objective： To evaluate the effect of hypoxia-inducible factor-1 alpha（HIF-1α） on angiogenesis in human prostate cancer cells. Methods ： Human prostate cancer cells of the line LNCaP were cultured and transfected by the recombinant plasmid pcDNA3. 1 （-）/HIF-1α containing the gene HIF-1α with the Lipofectamine 2000 system. The positive clone cells were selected by G418 and confirmed by Western blotting and immunofluorescence （ LNCaP/HIF-1α cells）. The expressions of VEGF, iNOS and Ang-2 were detected by Western blotting. Results： The expression of HIF-1α in the LNCaP/HIF-1α cells was distinctly higher than that in the LNCaP cells. Compared with the LNCaP cells, the expressions of VEGF and iNOS were up-regulated, whereas Ang-2 remained unchanged in the LNCaP/HIF-1α cells. Conclusion： The over-expression of HIF-1α can induce an increase in angiogenesis proteins and improve the angiogenesis potency of prostate cancer.
出处
《中华男科学杂志》
CAS
CSCD
2007年第11期988-991,共4页
National Journal of Andrology
