Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
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Expression of Caspase 3, Survivin, and p53 Protein in Urethane Induced Mouse Lung Carcinogenesis

Urethane으로 유발된 생쥐 폐샘암종 발생과정에서 Caspase 3, Survivin과 p53 단백 발현

  • Shin, Jong Wook (Division of Allergy, Respiratory and Critical Care Medicine, Department of Internal Medicine, Chung Ang University College of Medicine) ;
  • Lee, Soo Hwan (Department of Pathology, Chung Ang University College of Medicine) ;
  • Park, Eon Sub (Department of Pathology, Chung Ang University College of Medicine)
  • 신종욱 (중앙대학교 의과대학 내과학교실) ;
  • 이수환 (중앙대학교 의과대학 병리학교실) ;
  • 박언섭 (중앙대학교 의과대학 병리학교실)
  • Received : 2007.05.03
  • Accepted : 2007.09.14
  • Published : 2007.09.30
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Abstract

Purpose: An imbalance of cell proliferation and cell apoptosis is an important mechanism in carcinogenesis. Capase 3, survivin and p53 have been identified as important members of the apoptotic related proteins. This study evaluated the proliferating cell nuclear antigen(PCNA), apoptosis, apoptotic related protein such as capase 3, survivin and p53 using urethane-induced mouse lung carcinogenesis, which provides reproducible steps from hyperplasia to adenocarcinoma. Methods: Urethane was administered to the ICR mice through an intra-peritoneal injection, The mice were sacrificed at 5, 15, and 25 weeks after urethane intervention. The sequential morphological changes and immunohistochemical expression of PCNA, apoptosis, capase 3, survivin, and p53 were examined during mouse lung carcinogenesis. Results: During carcinogenesis, the sequential histological changes were observed from hyperplasia of type II pneumocytes, to anadenoma, and ultimately to an overt adenocarcinoma. The PCNA Labeling index (LI) was 9.6% in hyperplasia, 23.2% in adenoma, and 55.7% in adenocarcinoma, respectively. The apoptotic LI was 0.24% in hyperplasia, 1.25% in adenoma, and 5.27% in adenocarcinoma. A good correlation was observed between the PCNA LI and apoptotic LI. The expression of caspase 3 was remarkable- i.e., 46.7% in adenocarcinoma, in contrast to 15% in hyperplasia and 16% in adenoma. Survivin was detected weakly in the alveolar hyperplasia and showed an increasing expressional pattern in adenoma and adenocarcinoma. p53 expression was detected only in the adenocarcinoma lesions with an expression rate of 13.3%. The level of caspase 3 expression correlated with the increase in the apoptotic index. The positive expression of caspase 3 was associated with an increased apoptotic index. Conclusions: These results suggest that the PCNA LI and apoptotic LI might be useful markers for evaluating the risk of a malignant transformation. In addition, caspase, survivin and p53 might play a role in the early and late steges of urethane-induced mouse lung carcinogenesis.

연구배경 및 목적: ICR계의 생쥐에 Urethane을 투여하여 발생되는 폐 병변의 형태변화를 관찰하고, 폐 샘암종으로 진행과정에서 세포증식능과 세포자멸사, 세포자멸사와 관련된 조절인자인 caspase3, 자멸사 억제인자인 survivin, 그리고 종양억제 유전자 산물인 p53 단백질이 발암과정에서 발현되는 양상을 관찰함으로써 발암과정에서의 역할을 규명하고자 하였다. 방법: Urethane을 ICR 생쥐에게 복강 내 주사를 하였고, 5주, 15주, 25주에 폐병변을 육안적으로 헤마토실린 및 에오신 염색을 관찰하였고, 면역조직화학적 방법으로 PCNA지수, 세포자멸사 지수, capase 3, survivin 및 p53의 발현을 관찰하였다. 결과: Urethane 투여 5주부터 폐의 증식이 관찰되었고, 샘종은 10주 이후부터 출현하여 시간 경과에 따라 크기와 구조적 변화가 동반되었고, 세포학적 이상소견과 더불어 주변으로 침윤성 변화가 있는 샘암종은 25주 이후에 출현하였다. 세포증식능과 세포자멸사 지수는 폐의 증식증에서는 9.6%와 0.24%, 샘종에서는 23.2%와 1.25%, 그리고 샘암종에서는 55.7%와 5.21%이었다. 따라서 세포증식능과 세포자멸사는 폐 샘암종 발생초기부터 통계적으로 유의하게 지속적으로 증가하였고, 특히 샘암종으로 진행할 경우 현저하게 증가하였다. caspase 3는 증식증에서는 15%, 샘종은 16%의 발현율을 보이는 반면, 폐샘암종은 46.7%의 발현율을 보이면서 암 단계에서 현저하게 발현이 증가하였다. Survivin 단백의 발현은 폐의 증식증, 샘종, 그리고 샘암종으로 진행할수록 발현빈도가 통계적의로 유의하게 증가하였다. p53 단백은 폐의 증식증과 샘종에서는 전혀 발현되지 않았으나 침윤성 샘암종의 일부에서 발현되었다. 이상의 결과로 생쥐의 폐샘암종 발생과정에서 세포증식능과 세포자멸사는 종양의 발생과 진행과정에서 지속적으로 관여함을 알 수 있었다. 또한 survivin 발현은 샘암종의 초기단계에서부터 지속적으로 관여하며, p53 유전자 변이는 초기보다는 암종으로 형질전환이 일어난 후에 부분적으로 발생하는 것으로 보인다. 결론: 본 연구를 통하여 PCNA와 세포자멸사 지수는 암세포로 형질전환하는 위험도를 평가하는 지표로 유용할 것으로 보이며, caspase, survivin과 p53는 urethane에 의해 유도된 생쥐 폐암 모델에서 발암과정에 중요하게 관여하는 단백질로 보인다.

Keywords

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