AKAP12基因启动子甲基化在哈萨克族食管鳞癌中研究

摘要

Objective To explore the promoter methylation of AKAP12 gene in patients with esophageal squamous cell carcinoma( ES-CC) , and its relationship with clinical parameters. Methods AKAP12 methylation status was detected in Eca109 treated by 5-aza-CdR or normal culture, and 18 pairs of Kazakh’ s ESCC tissues by methylation specific PCR( MSP) . The expression of AKAP12 in ESCC cell lines Eca109 and 18 pairs of Kazakh ESCC tissues was detected by real time PCR(RT-PCR). Results The methylation rate of AKAP12 and the normal cells was 0 in 5-aza-CdR-treated Eca109 cells and 5. 6%(1/18) in ESCC tissues. The expression of AKAP12 in 5-aza-CdR group was 0. 99 times as high as control group withβ-actin as the internal reference(P=0. 96), 1. 93 times with GAPDH as the internal reference(P=0. 47). The expression of AKAP12 in the tumor tissues of 18 pairs Kazakh’s ESCC was 0. 86 times as high as that of paracancerous tissues withβ-actin as the internal reference(P=0. 74),2. 87 times with GAPDH as the internal reference(P=0. 11). The expression and the methylation rate of AKAP12 were not correlated with sex, morphologic type, differentiation and metasta-ses(P>0. 05). Conclusion The methylation of AKAP12 is relatively low in esophageal squamous cell carcinoma.%目的：研究A激酶锚定蛋白12(A kinase anchoring protein 12,AKAP12)基因在哈萨克族食管鳞癌(esophageal squa-mous cell carcinoma,ESCC)中,是否发生高度甲基化及其与临床病理参数间的关系。方法应用甲基化特异性PCR( MSP)检测用5-aza-CdR处理及正常培养的食管鳞癌细胞系Eca109和18对哈萨克族食管鳞癌组织中AKAP12的甲基化状态。通过实时荧光定量PCR( RT-PCR)检测食管鳞癌细胞系Eca109和18对哈萨克族食管鳞癌组织中AKAP12的表达量。结果AKAP12在Eca109细胞系,5-aza-CdR处理组及正常培养组中甲基化率为0；在18对哈萨克族食管鳞癌组织中甲基化率为5.6%(1/18)。 AKAP12在5-aza-CdR组表达量与对照组以β-actin为内参相差0.99倍(P=0.96),以GAPDH为内参相差1.93倍(P=0.47)；在18对哈族食管鳞癌组织中,癌组织与癌旁组织表达量以β-actin为内参相差0.86倍(P=0.74),以GAPDH为内参相差2.87倍(P=0.11)。 AKAP12表达量及甲基化率与性别、大体类型、分化程度及有无转移无相关性(P>0.05)。