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目的 :探讨缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)与着丝粒蛋白W(centromere protein W,CENP-W)在人胶质瘤中表达的相关性。方法 :采用免疫组织化学法分别检测高级别胶质瘤、低级别胶质瘤及瘤旁正常脑组织中HIF-1α和CENP-W的表达水平,并统计学分析蛋白表达水平与胶质瘤临床病理特征的关系,以及两蛋白表达之间的相关性。用氯化钴抑制胶质瘤U251细胞中HIF-1α的分解,然后分别采用蛋白质印迹法和实时荧光定量PCR法检测CENP-W蛋白及m RNA表达的变化。应用特异性小干扰RNA(small interference RNA,si RNA)技术下调U251细胞的HIF-1α表达后,采用蛋白质印迹法和实时荧光定量PCR法检测CENP-W蛋白及m RNA表达的变化。应用特异性si RNA下调U251细胞中CENP-W表达后,采用蛋白质印迹法和实时荧光定量PCR法检测HIF-1α蛋白及m RNA表达的变化。结果:胶质瘤组织中HIF-1α和CENP-W的表达水平明显高于癌旁正常组织(P<0.05),并且与胶质瘤的病理级别明显相关(P<0.05);两蛋白表达水平之间也呈正相关性(r=0.709,P<0.05)。氯化钴作用U251细胞后,HIF-1a和CENP-W蛋白及m RNA的表达水平均明显升高(P<0.01)。si RNA干扰HIF-1α表达后,CENP-W蛋白及m RNA表达水平均明显降低(P<0.01);而si RNA干扰CENP-W表达后,HIF-1α蛋白及m RNA表达水平无明显变化(P>0.05)。结论:HIF-1α和CENP-W表达均与胶质瘤的病理级别呈正相关性。HIF-1α可以调节CENP-W的表达。
Objective: To investigate the correlation between hypoxia-inducible factor-1α (HIF-1α) and centromere protein W (CENP-W) expression in human glioma. Methods: The expression of HIF-1α and CENP-W in high grade gliomas, low grade gliomas and normal parahippocampal tissues were detected by immunohistochemical method. The expression of HIF-1α and CENP-W in high grade gliomas, The relationship between the pathological features, and the correlation between the two protein expressions. Inhibition of HIF-1αin human umbilical vein glioma U251 cells was induced by cobalt chloride. The changes of CENP-W protein and m RNA expression were detected by Western blotting and real-time fluorescence quantitative PCR respectively. After down-regulating the expression of HIF-1α in U251 cells by siRNA, the changes of CENP-W protein and m RNA expression were detected by Western blotting and real-time fluorescence quantitative PCR. The expression of CENP-W in U251 cells was down-regulated by specific si RNA. The expression of HIF-1α protein and m RNA were detected by Western blotting and real-time fluorescence quantitative PCR. Results: The expression of HIF-1α and CENP-W in glioma tissues was significantly higher than that in adjacent normal tissues (P <0.05), and significantly correlated with the pathological grade of gliomas (P <0.05) Also had a positive correlation (r = 0.709, P <0.05). The expression of HIF-1a, CENP-W protein and m RNA in U251 cells were significantly increased after cobalt chloride treatment (P <0.01). The expression of CENP-W protein and m RNA were significantly decreased after si RNA interference with HIF-1α expression (P <0.01). However, the mRNA and protein expression of HIF-1α did not change significantly after si RNA interference with CENP-W P> 0.05). Conclusion: The expression of HIF-1α and CENP-W are positively correlated with the pathological grade of gliomas. HIF-1α regulates CENP-W expression.