Hpall-Mspl Methylation Microarray를 이용한 비소세포폐암의 DNA Methylation Marker 발굴

Identification of DNA Methylation Markers for NSCLC Using Hpall-Mspl Methylation Microarray

  • 권미혜 (건양대학교병원 호흡기내과) ;
  • 이고은 (건양대학교병원 호흡기내과) ;
  • 권선중 (건양대학교병원 호흡기내과) ;
  • 최유진 (건양대학교병원 호흡기내과) ;
  • 나문준 (건양대학교병원 호흡기내과) ;
  • 조현민 (건양대학교병원 흉부외과) ;
  • 김영진 (건양대학교병원 흉부외과) ;
  • 설혜정 (건양대학교병원 병리학과) ;
  • 조영준 (건양대학교병원 방사선학과) ;
  • 손지웅 (건양대학교병원 호흡기내과)
  • Kwon, Mi Hye (Departments of Internal Medicine, Konyang University School of Medicine) ;
  • Lee, Go Eun (Departments of Internal Medicine, Konyang University School of Medicine) ;
  • Kwon, Sun Jung (Departments of Internal Medicine, Konyang University School of Medicine) ;
  • Choi, Eugene (Departments of Internal Medicine, Konyang University School of Medicine) ;
  • Na, Moon Jun (Departments of Internal Medicine, Konyang University School of Medicine) ;
  • Cho, Hyun Min (Departments of Chest Surgery, Konyang University School of Medicine) ;
  • Kim, Young Jin (Departments of Chest Surgery, Konyang University School of Medicine) ;
  • Sul, Hye Jung (Departments of Pathology, Konyang University School of Medicine) ;
  • Cho, Young Jun (Departments of Radiology, Konyang University School of Medicine) ;
  • Son, Ji Woong (Departments of Internal Medicine, Konyang University School of Medicine)
  • 투고 : 2008.10.09
  • 심사 : 2008.11.19
  • 발행 : 2008.12.30

초록

연구배경: 유전자의 후생적인 변화(epigenetic alteration)는 악성종양의 병인론에 있어서 유전자 변이와 동등한 위치를 점하고 있다. 특히 종양억제 유전자의 전사 촉진(promoter) 부위에 발생하는 비정상적인 메칠화(methylation)는 유전자의 발현을 침묵화(silencing)하고, 결과적으로 유전자의 기능 소실을 일으키게 된다. 저자들은 CpG island와 HpaII site를 가지고 있으며 암화 과정에 관여할 것으로 생각되는 유전자에 대하여 HpaII-MspI methylation microarray를 이용하여 새로운 종양억제 유전자를 발굴하고자 하였다. 방 법: 2005년 건양대학교 병원에서 수술한 비 소세포성 폐암 환자 10명에서 폐암조직과 상응하는 암 주변의 정상조직을 얻었으며, HpaII-MspI methylation microarray (Methyl-Scan DNA chip$^{(R)}$, Genomic tree, Inc, South Korea)를 이용하여 21개의 유전자에 대하여 DNA methylation profile을 분석하였다. 각각의 유전자에서 메칠화된 정도를 두 그룹에서 비교하였고, 정상 대조군으로 두 명의 젊고 건강한 기흉 환자에서 수술한 폐 조직에 대하여 methylation profile을 분석하였다. 결 과: 21개의 대상 유전자 중 10개의 유전자에서 폐암조직, 폐암 주변 정상 조직, 대조군에서 모두 공통적으로 과메칠화 되었고, 나머지 11개의 유전자 중 APC, AR, RAR-b, HTR1B, EPHA3, CFTR의 6개의 유전자에서 대조군에서 메칠화가 없으며, 폐암조직에서 폐암 주변 정상 조직에 비하여 더 빈번하게 과메칠화 되었다. 결 론: HTR1B, EPHA3, CFTR은 비소세포 폐암에서 후생적 변화로 발생하는 새로운 종양억제 유전자의 후보 유전자로서의 가능성이 있을 것으로 생각한다.

Background: Epigenetic alterations in certain genes are now known as at least important as genetic mutation in pathogenesis of cancer. Especially abnormal hypermethylation in or near promoter region of tumor suppressor genes (TSGs) are known to result in gene silencing and loss of gene function eventually. The authors tried to search for new lung cancer-specific TSGs which have CpG islands and HpaII sites, and are thought to be involved in carcinogenesis by epigenetic mechanism. Methods: Tumor tissue and corresponding adjacent normal tissue were obtained from 10 patients who diagnosed with non small cell lung cancer (NSCLC) and underwent surgery in Konyang university hospital in 2005. Methylation profiles of promoter region of 21 genes in tumor tissue & non-tumor tissue were examined with HpaII-MspI methylation microarray (Methyl-Scan DNA chip$^{(R)}$, Genomic tree, Inc, South Korea). The rates of hypermethylation were compared in tumor and non-tumor group, and as a normal control, we obtained lung tissue from two young patients with pneumothorax during bullectomies, methylation profiles were examined in the same way. Results: Among the 21 genes, 10 genes were commonly methylated in tumor, non-tumor, and control group. The 6 genes of APC, AR, RAR-b, HTR1B, EPHA3, and CFTR, among the rest of 11 genes were not methylated in control, and more frequently hypermethylated in tumor tissue than non-tumor tissue. Conclusion: In the present study, HTR1B, EPHA3, and CFTR are suggested as possible novel TSGs of NSCLC by epigenetic mechanism.

키워드

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