【摘要】 目的:探讨在乏氧条件下,HIF-1α、VEGF及CXCR4在鼻咽癌细胞中的表达,并探讨HIF-1α与VEGF和CXCR4的关系。方法:利用荧光定量PCR方法检测不同乏氧时相HIF-1α、VEGF及CXCR4 mRNA表达。利用Western bolt检测HIF-1α蛋白表达。构建特异性的HIF-1α慢病毒干扰载体,利用慢病毒感染鼻咽癌细胞,杀稻瘟菌素进行药物筛选,利用荧光定量PCR和Western blot检测干扰效果。利用荧光定量PCR检测HIF-1α表达沉默后,鼻咽癌细胞VEGF和CXCR4的表达变化。结果:在乏氧条件下,鼻咽癌细胞CNE2HIF-1α mRNA表达稳定,蛋白表达增强,VEGF及CXCR4的mRNA表达均明显增强。利用特异性的HIF-1α慢病毒干扰鼻咽癌细胞CNE2,CNE2细胞中HIF-1α表达降低了81%,说明HIF-1α载体构建成功。HIF-1α表达沉默后,VEGF及CXCR4的表达水平也明显降低。结论:乏氧可以诱导鼻咽癌细胞HIF-1α、VEGF和CXCR4的表达,且VEGF和CXCR4的表达受HIF-1α的表达调控。更多还原

【Abstract】 OBJECTIVE: To study the expressions of hypoxia inducible factor-1α (HIF-1α), chemokine receptor CXCR4 and vascular endothelial growth factor (VEGF) in nasopharyngeal carcinoma under hypoxia and to explore the relationship among them.METHODS: The expressions of HIF1α, CXCR4 and VEGF at different hypoxic culture phases were detected by Real Time PCR or by Western blot. HIF1α specific RNAi lentiviral vector was constructed.HIF-1α silencing cells were screened by blasticidin.Real Time PCR and Western blot analysis were used to detect the expressions of HIF-1α mRNA and protein, respectively.Real Time PCR was used to observe the changes of VEGF and CXCR4 expressions after HIF-1α silencing. RESULTS: Under hypoxia, the mRNA expressions of VEGF and CXCR4 were upregulated, while HIF-1α mRNA expression was not obviously altered. HIF-1α protein expression increased under hypoxia. Transduction of nasopharyngeal carcinoma cell with HIF-1α RNAi vector resulted in sequence specific silencing with 81% decreases of HIF-1α expression. The expressions of VEGF and CXCR4 decreased after HIF-1α silencing.CONCLUSIONS: Hypoxia could induce HIF-1α, VEGF and CXCR4 expressions. HIF-1α silencing could downregulate VEGF and CXCR4 expressions.更多还原