CAJ | 학술논문

目的探讨人食管癌相关基因4(ECRG4)在食管癌中表达缺失的机制.方法采用聚合酶链反应-单链构象多态(PCR-SSCP)和DNA测序的方法检测80对配对食管鳞状细胞癌(ESCC)肿瘤组织和癌旁正常上皮中ECRG4的外显子突变;采用DNA亚硫酸氧盐修饰和序列特异性聚合酶链反应(ssPCR)检测EC9706细胞系ECRG4基因启动子CpG岛甲基化状态;采用逆转录聚合酶链反应(RT-PCR)检测去甲基化药物5-氮杂-2-脱氧胞苷或三氧化二砷(As2O3)处理后ECRG4 mRNA的重新表达.结果 80对ESCC配对标本中,ECRG4的4个外显子编码区均未发现突变.EC9706细胞系ECRG4基因核心启动子区16个CpG岛中,有11个呈高甲基化状态,ECRG4 mRNA不表达.EC9706细胞处理前ECRG4 mRNA不表达,去甲基化药物处理后,ECRG4 mRNA均重新表达.结论甲基化表观遗传学机制是导致ECRG4基因在食管癌细胞系EC9706中表达缺失的一个机制.
목적 탐토인식관암상관기인4(ECRG4)재식관암중표체결실적궤제.방법 채용취 합매련반응-단련구상다태(PCR-SSCP)화DNA측서적방법검측80대배대식관린상세포암(ESCC)종류조직화암방정상상피중ECRG4적외현자돌변;채용DNA아류산양염수식화서렬특이성취합매련반응(ssPCR)검측EC9706세포계ECRG4기인계동자CpG도갑기화상태;채용역전록취합매련반응(RT-PCR)검측거갑기화약물5-담잡-2-탈양포감혹삼양화이신(As2O3)처리후ECRG4 mRNA적중신표체.결과 80대ESCC배대표본중,ECRG4적4개외현자편마구균미발현돌변.EC9706세포계ECRG4기인핵심계동자구16개CpG도중,유11개정고갑기화상태,ECRG4 mRNA불표체.EC9706세포처리전ECRG4 mRNA불표체,거갑기화약물처리후,ECRG4 mRNA균중신표체.결론 갑기화표관유전학궤제시도치ECRG4기인재식관암세포계EC9706중표체결실적일개궤제.
Objective To investigate the mechanism of loss of human esophageal cancer-related gene 4 (ECRG4) expression in esophageal squamous cell carcinoma (ESCC.) Methods PCR-SSCP and DNA sequencing analysis were used to detect the mutation of ECRG4 exons in esophageal cancer and matched adjacent normal tissues of 80 patients. DNA bisulfite-modifying ssPCR sequencing assay was used to examine the methylation status of ECRG4 promoter in human esophageal squamous cell carcinoma EC9706 cells. The re-expression of ECRG4 mRNA was examined by RT-PCR in EC9706 cells, after treatment with either demethylation drug 5-aza-2'-deoxycytidine or arsenic trioxide. Results No mutation in the four ECRG4 exons was found in all the ESCC and matched normal adjacent tissues. RT-PCR showed that 11 of 16 CpG islands of ECRG4 promoter were hypermethylated, while ECRG4 mRNA expression level was undetectable in the EC9706 cells. The ECRG4 mRNA was re-expressed after treatment with either demethylation drug 5-aza-2'-deoxycytidine or arsenic trioxide. Conclusion The epigenetic mechanism of methylation is a reason of loss of ECRG4 gene expression in the ESCC cell line EC9706.