HOXA11基因启动子区甲基化是胰腺癌的潜在诊断标志物

Methylation of HOXA11 is a potential diagnostic marker of pancreatic cancer

目的在胰腺癌中探讨HOXA11基因启动子区甲基化状态及其作为诊断标志物的可能性。方法应用半定量RT-PCR和甲基化特异性PCR(methylation-specific PCR,MSP)技术检测8株胰腺癌细胞系中HOXA11的表达及启动子区甲基化情况,应用MSP在216例胰腺癌组织中检测HOXA11启动子区甲基化情况,并分析HOXA11启动子区甲基化与临床病理因素之间的相关性。结果HOXA11在Panc3.11、Panc5.04、Panc10.05、BXPC3细胞中表达缺失,其MSP结果为完全甲基化。在SW1990、AsPC1中高表达,其MSP结果为非甲基化,在CFPAC1和MIAPaCa2细胞中表达减低,其MSP结果呈部分甲基化状态。应用5-氮杂-2′-脱氧胞苷(5-aza-2′-deoxycytidine,5-aza)处理细胞后,HOXA11在Panc3.11、Panc5.04、Panc10.05、BXPC3细胞中恢复表达,在CFPAC1和MIAPaCa2细胞中表达增加,在SW1990、AsPC1细胞中表达水平无明显变化。在216例原发性胰腺癌患者中,76.85%(166/216)发生HOXA11甲基化,甲基化与肿瘤分化程度相关(P<0.0003)。结论HOXA11基因在胰腺癌组织中频繁发生甲基化,其表达受启动子区甲基化调控,HOXA11甲基化是胰腺癌潜在的诊断标志物。

Objective To investigate the methylation status of HOXA11 gene in human pancreatic cancer and its potential application in the diagnosis.Methods Semi-quantitative RT-PCR and methylation-specific PCR(MSP)were used to detect HOXA11 expression and promoter region methylation in 8 pancreatic cancer cell lines and 216 cases of primary pancreatic cancer samples.Chi-square test was used to analyze the association of HOXA11 methylation and clinical pathological factors in pancreatic cancer.Results The expression of HOXA11 was lost in Panc3.11,Panc5.04,Panc10.05 and BXPC3,and the promoter region of HOXA11 was completely methylated in these cells.The expression of HOXA11 gene was reduced in CFPAC1 and MIAPaCa2 cells,and its promoter region was partially methylated.It was highly expressed and the promoter region was unmethylated in SW1990 and AsPC1 cells.The expression of HOXA11 was unchanged in SW1990 and AsPC1 cells,increased in CFPAC1 and MIAPaCa2 cells,and restored in Panc3.11,Panc5.04,Panc10.05,BXPC3 cells after 5-aza treatment.HOXA11 was methylated in 76.85%(166/216)of primary pancreatic cancer,and methylation of HOXA11 was associated with tumor differentiation(P<0.0003).Conclusion HOXA11 gene is frequently methylated in pancreatic cancer and its expression is regulated by promoter methylation.HOXA11 methylation is a potential diagnostic marker in pancreatic cancer.