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目的:探讨唾液腺腺样囊性癌(adenoid cystic carcinoma,ACC)细胞系中CDH13、RASSF2A、TFPI-2、hMLH-1、MyoD1基因的甲基化及其与mRNA表达之间的关系。方法:甲基化特异性PCR(methylation-specific PCR,MSP)法检测ACC细胞系ACC-2、ACC-3、ACC-M中CDH13、RASSF2A、TFPI-2、hMLH-1、MyoD1基因的甲基化;RT-PCR及荧光实时定量PCR对存在甲基化的基因进行mRNA水平的检测。结果:3个细胞系中均存在CDH13、RASSF2A、TFPI-2基因的甲基化和未甲基化,只存在MyoD1基因的甲基化和hMLH-1基因的未甲基化;ACC-2中CDH13基因mRNA水平的表达显著高于ACC-M,ACC-3中RASSF2A的表达显著高于ACC-2、ACC-M,3个细胞系中均未检测到MyoD1基因的表达。结论:ACC细胞系中,CDH13、RASSF2A、TFPI-2、MyoD1基因的甲基化为常见事件,甲基化可能为MyoD1基因失活的主要机制,CDH13基因的表达可能与ACC的转移相关。
Objective: To investigate the methylation of CDH13, RASSF2A, TFPI-2, hMLH-1 and MyoD1 in adenoid cystic carcinoma (ACC) cell lines and its relationship with mRNA expression. Methods: The methylation-specific PCR (MSP) was used to detect the methylation of CDH13, RASSF2A, TFPI-2, hMLH-1 and MyoD1 genes in ACC-2, ACC-3 and ACC- ; RT-PCR and real-time quantitative PCR of mRNA methylation exists for the detection of mRNA levels. Results: The methylation and unmethylation of CDH13, RASSF2A and TFPI-2 genes existed in all three cell lines, only methylation of MyoD1 gene and unmethylation of hMLH-1 gene existed in ACC-2 The expression of CDH13 mRNA was significantly higher than that of ACC-M. The expression of RASSF2A in ACC-3 was significantly higher than that in ACC-2 and ACC-M. No expression of MyoD1 was found in all three cell lines. CONCLUSION: Methylation of CDH13, RASSF2A, TFPI-2 and MyoD1 genes in ACC cell lines is a common event. Methylation may be the main mechanism of MyoD1 gene inactivation. CDH13 gene expression may be related to ACC metastasis.