HIF-1α基因表达沉默对鼻咽癌组织VEGF和CXCR4表达影响的研究  

Effect of silencing HIF-1α on expressions of VEGF and CXCR4 in nasopharyngeal carcinoma cell line

作　　者：吴德华[1] 朱晓霞[1] 张洪波[1] 李高峰[1] 陈龙华[1] 

机构地区：[1]南方医科大学南方医院放疗科,广东广州510515

出　　处：《中华肿瘤防治杂志》2008年第3期170-173,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30500242;30672312);广东省自然科学基金(05004743)~~

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。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 a- mong them. METHODS.. The expressions of HIF1α, CXCR4 and VEGF at different hypoxic culture phases were detected by Real Time PCR or by Western blot. HIF1a 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.

关 键 词：鼻咽肿瘤 基因 RNA干扰 

分 类 号：R739.63[医药卫生—肿瘤]