胰腺癌细胞株HOXA7基因表达及其启动子区甲基化状态

目的 检测HOXA7 mRNA在胰腺癌细胞株中的表达及其启动子区的甲基化状态,探讨两者的相关性.方法 采用RT-PCR法检测人胰腺癌细胞株BxPC3、CFPAC1、PANC1和SW1990细胞的HOXA7 mRNA的表达水平.采用重亚硫酸盐测序PCR(bisulfite sequencing PCR,BSP)和结合重亚硫酸盐的限制性内切酶法(combined bisulfite restriction analysis,COBRA)检测启动子区域甲基化状态.应用去甲基化药物5-氮杂-2'-脱氧胞苷(5-aza-2-deoxycytidine,5-aza-dC)处理各细胞株,检测处理前后细胞HOXA7 mRNA表达和甲基化状态的变化.结果 胰腺癌细胞株BxPC3、CFPAC1和SW1990细胞均表达HOXA7 mRNA,而PANC1细胞不表达HOXA7 mRNA.CFPAC1、BxPC3、PANC1和SW1990中HOXA7启动子甲基化率分别为93.16%、90.65%、90.09%、52.30%,SW1990细胞的HOXA7甲基化率较其他三株细胞均明显降低(P值均＜0.01).经5-aza-dC处理后,PANC1细胞的HOXA7 mRNA重新表达,BxPC3的HOXA7 mRNA表达增强;而CFPAC1和SW1990细胞的HOXA7 mRNA在5-aza-dC处理前后无明显变化.结论 胰腺癌细胞株BxPC3和PANC1细胞的HOXA7mRNA表达与启动子区甲基化状态密切相关,而CFPAC 1和SW1990细胞两者无明显相关性.

Expression and promoter methylation status of HOXA7 gene in pancreatic cancer cell line

Objective To investigate the expression and methylation status of HOXA7 gene in human pancreatic cancer cell lines, and to explore the relationship between them.Methods HOXA7 mRNA expression of human pancreatic cancer cell lines BxPC3, CFPAC1, PANC1 and SW1990was detected by RT PCR.Bisulfite sequencing PCR (BSP) and combined bisulfite restriction analysis (COBRA) was used to test promoter methylation status.All the cell lines were treated by 5-aza-2-deoxycytidine (5-aza-dC), and HOXA7mRNA expression, methylation status was detected before and after this treatment.Results HOXA7 mRNA was expressed in BxPC3, CFPAC1 and SW1990, while there was no expression of HOXA7 mRNA in PANC1.HOXA7 promoter methylation rates of CFPAC1, BxPC3, PANC1 and SW1990 were 93.16%, 90.65%,90.09% ,52.30%.HOXA7 promoter methylation rate of SW1990 was significantly lower than those in other 3cell lines ( P ＜0.01 ).After 5-aza-dC treatment, HOXA7 mRNA of PANC1 was expressed again, and HOXA7mRNA of BxPC3 was increasingly expressed;while the expression of HOXA7 mRNA in CFPAC1 and SW1990was not significantly changed after 5-aza-dC treatment.Conclusions The expression of HOXA7 mRNA in BxPC3 and PANC1 was closely correlated with promoter hypermethylation, while there was no obvious relation in CFPAC 1 and SW1990.