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目的:研究NF-κB对人非霍奇金淋巴瘤(NHL)HIF-1α-VEGF途径的调控及其机制。方法:应用Echinomycin(EC)处理人NHL细胞,将HIF-1α反义质粒转染至肿瘤细胞中,采用蛋白质印迹法检测经两种方法处理后各细胞系中VEGF表达水平。以NF-κB特异性抑制剂quinazoline(QNZ)及Bay11-7082预处理人NHL细胞,检测各细胞系中HIF-1α蛋白的表达水平,同时采用实时定量PCR方法检测HIF-1αmRNA变化。应用QNZ处理NHL细胞后检测HIF-1α下游调控靶点VEGF的蛋白表达水平。结果:通过应用HIF-1α特异性抑制剂和转染反义质粒两种方法均能够抑制HIF-1α,明显下调了VEGF蛋白表达水平,与对照组相比差异均有统计学意义,P<0.05。NF-κB抑制剂QNZ及Bay11-7082能够降低NHL细胞HIF-1α蛋白及基因水平的表达,同时下调其下游调控靶点VEGF蛋白的表达,P<0.05。结论:抑制NF-κB可阻断人NHL细胞HIF-1α-VEGF通路,其机制可能与NF-κB调控HIF-1α的基因与蛋白表达有关。
Objective: To investigate the regulation of NF-κB on HIF-1α-VEGF pathway in human non-Hodgkin’s lymphoma (NHL) and its mechanism. Methods: Human NHL cells were treated with Echinomycin (EC). The antisense plasmid of HIF-1α was transfected into tumor cells. The expression of VEGF in each cell line was detected by Western blotting. The human NHL cells were pretreated with quinazoline (QNZ) and Bay11-7082, which are specific inhibitors of NF-κB, to detect the expression of HIF-1αprotein in each cell line, and the changes of HIF-1αmRNA were detected by real-time quantitative PCR. After treatment of NHL cells with QNZ, the protein expression of VEGF downstream of HIF-1α was detected. Results: Both HIF-1α-specific inhibitor and transfection antisense plasmid could inhibit HIF-1α expression and down-regulate the expression of VEGF protein significantly compared with the control group (P <0.05) . NF-κB inhibitors QNZ and Bay11-7082 could reduce the expression of HIF-1αprotein and gene in NHL cells, and down-regulate the expression of VEGF protein downstream of it, P <0.05. CONCLUSION: Inhibition of NF-κB can block the HIF-1α-VEGF pathway in human NHL cells, which may be related to the regulation of gene and protein expression of HIF-1α by NF-κB.