Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
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Microsatellite Alterations of Plasma DNA in Non Small Cell Lung Cancer

비소세포폐암 환자의 혈장 DNA를 이용한 Microsatellite 분석

  • Kim, Kyu-Sik (Lung and Esophageal Cancer Clinic, Chonnam National University Hwasun Hospital) ;
  • Kim, Eun-Jung (Department of Pulmonology and Critical Care Medicine, Chonnam National University Medical School) ;
  • Kim, Soo-Ock (Department of Pulmonology and Critical Care Medicine, Chonnam National University Medical School) ;
  • Oh, In-Jae (Department of Pulmonology and Critical Care Medicine, Chonnam National University Medical School) ;
  • Park, Chang-Min (Department of Pulmonology and Critical Care Medicine, Chonnam National University Medical School) ;
  • Jeong, Ju-Yeon (Lung and Esophageal Cancer Clinic, Chonnam National University Hwasun Hospital) ;
  • Kim, Yu-Il (Department of Pulmonology and Critical Care Medicine, Chonnam National University Medical School) ;
  • Lim, Sung-Chul (Department of Pulmonology and Critical Care Medicine, Chonnam National University Medical School) ;
  • Park, Jong-Tae (Department of Forensic Medicine, Chonnam National University Medical School) ;
  • Kim, Young-Chul (Lung and Esophageal Cancer Clinic, Chonnam National University Hwasun Hospital)
  • 김규식 (화순 전남대학교 병원 폐식도 종양 클리닉) ;
  • 김은정 (전남대학교 의과대학 호흡기 내과학교실) ;
  • 김수옥 (전남대학교 의과대학 호흡기 내과학교실) ;
  • 오인재 (전남대학교 의과대학 호흡기 내과학교실) ;
  • 박창민 (전남대학교 의과대학 호흡기 내과학교실) ;
  • 정주연 (화순 전남대학교 병원 폐식도 종양 클리닉) ;
  • 김유일 (전남대학교 의과대학 호흡기 내과학교실) ;
  • 임성철 (전남대학교 의과대학 호흡기 내과학교실) ;
  • 박종태 (전남대학교 의과대학 법의학교실) ;
  • 김영철 (화순 전남대학교 병원 폐식도 종양 클리닉)
  • Received : 2004.12.20
  • Accepted : 2005.01.20
  • Published : 2005.04.30
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Abstract

Microsatellites are short tandem repeated nucleotide sequences that are present throughout the human genome. Variations in the repeat number or a loss of heterozygosity around the microsatellites have been termed a microsatellite alteration (MA). A MA reflects the genetic instability caused by an impairment in the DNA mismatch repair system and is suggested to be a novel tumorigenic mechanism. A number of studies have reported that MA in the DNA extracted from the plasma occurs at varying frequencies among patients with a non-small cell lung carcinoma (NSCLC). The genomic DNA from 9 subjects with a non-small cell lung cancer (squamous cell cancer 6, adenocarcinoma 2, non-small cell lung cancer1) and 9 age matched non-cancer control subjects (AMC: tuberculosis 3, other inflammatory lung disease 6) and 12 normal control subjects (NC) were extracted from the peripheral blood leukocytes and plasma. Three microsatellite loci were amplified with the primers targeting the Gene Bank sequence D21S1245, D3S1300, and D3S1234. MA in the form of an allelic loss or a band shift was examined with 6% polyacrylamide gel electrophoresis and silver staining. None (0/12) of the NC subjects less than 40 years of age showed a MA in any of the three markers, while 88.9%(8/9) of the AMC above 40 showed a MA in at least one of the three markers (p<0.05). Sixty percent(6/10) of the control subjects with a smoking history showed a MA in one of the three markers, while 9.1%(1/11) of the control subjects without smoking history showed a MA (p<0.05). However, not only did 66.7%(6/9) of lung cancer patients show a MA in at least one of the three markers but so did 88.9%(8/21) of the AMC patients (p>0.05). In conclusion, a MA in the D21S1245, D3S1300, and D3S1234 loci using DNA extracted from the plasma was detected in 66.7% of lung cancer while no MA was found in the young non-smoking control subjects. However, many of the non-cancer control subjects (aged smokers) also showed a MA, which compromised the specificity of the MA analysis as a screening test. Therefore, a further study with a larger sample size will be needed.

폐암의 조기 진단을 위한 방법으로써 MA의 의의를 알아보고자 전남대학교 병원 내과에 내원한 폐암 환자 9례(squamous cell carcinoma 6례, adenocarcinoma 2례, non-small cell lung cancer: 1례), 연령이 비슷한 비폐암 대조군 9례(AMC, 결핵: 3례, 비특이적 염증성 폐질환: 6례)와 40세 이하 정상인 12례(NC)를 대상으로, 이들의 말초혈액의 백혈구와 혈장으로부터 DNA를 추출하여 D21S1245, D3S1300, D3S1234 유전자좌의 MA를 분석하였다. 세가지 유전자좌 중 어느 한 유전자좌에서라도 MA가 관찰되면 MA가 있는 것으로 인정하였다. MA는 NC에서는 관찰되지 않았으나 0%(0/12), AMC에서는 88.9%(8/9)에서 관찰되었다. AMC와 NC 총 21례 중 흡연자에서 70%(7/10) 비흡연자에서 9.1%(1/11) MA가 관찰되었다(p<0.05). 폐암군과 AMC 총 18례 중 AMC에서 88.9%(8/9), 폐암군에서 66.7%(6/9)를 보여 양군간에 서로 차이 없이 모두 높은 빈도로 관찰되었다(p>0.05). 결과적으로 혈장 DNA의 MA는 40세 이하의 정상인들에서는 발견되지 않으며 폐암 환자들에서 높은 빈도로 발견되었다. 그러나 고령의 흡연자들인 비폐암 대조 군에서도 높은 빈도로 MA가 관찰되므로 폐암 조기진단의 지표로써는 적합하지 않을 것으로 예상된다. 그러나 본 연구는 소수의 한정된 대상을 이용한 결과로써 다양한 연령층과 흡연력 그리고 조직형에 따라 세분화된 더 큰 대상 군을 이용한 연구가 추구되어야 할 것이다.

Keywords

References

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