Diethylnitrosamine으로 유도된 백서 간암 발생 과정 중 ErbB 수용체 단백들과 TGF-α의 발현 양상
Expression of ErbB Receptor Proteins and TGF-α during Diethylnitrosamine-induced Hepatocarcinogenesis in the Rat Liver
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목적: ErbB 수용체 단백들은 막횡단 티로신 키나제 수용체이며 그 리간드와 결합하면 활성화되어 다양한 생물학 반응을 나타내는데 특히 개체 발생과 발암에 관여한다고 알려져 있다. 저자들은 인체 간질환과 유사한 간경변과 간세포종양을 만드는 실험 동물 모델을 확립한 후 ErbB 수용체 단백들과 대표적인 리간드인 TGF-α의 발현을 연구하였다. 대상과 방법: 매주 한 번씩 DEN (50 mg/kg)을 53마리의 수컷 Wistar 백서의 복강 내에 18주 동안 주입하였다. 8주까지는 2주마다 2마리를 안락사시켰고, 10주부터 18주까지는 매주 5마리를 안락사시켰다. 육안으로 색깔과 형태가 이상한 간표면의 결절들을 크기에 따라 3군(N1; 3 mm x<5 mm, N2; 5 mm x<10 mm, N3; x 10 mm, 장경)으로 분류하였다. 종양 및 비종양 조직들에 대하여 조직검사를 같이 하였다. GSTp, TGF-α, EGF-R, ErbB2, ErbB3 및 ErbB에 대한 항체를 이용하여 면역조직화학 연구를 시행하였다. 결과: 10주째에 간섬유화가 시작되었고, 12주째에 뚜렷한 간경변증이 시작되었다. 전종양 병소, 간세포종 및 간세포암종은 각각 2주째, 11주째 그리고 15주째부터 관찰하였다. 간결절들 (N1: 397, N2: 258, N3: 64)은 조직에서 재생결절 (N1; 72.3%, N2, 15.9%, N3: 0%), 간세포종 (N1; 27.2%, N2; 82.2%, N3; 7.6%) 그리고 간세포암종 (N1; 0.5%, N2; 1.9%, N3; 92.4%)을 나타내었다. EGF-R 발현은 12.5%의 재생결절, 64.7%의 간세포종 그리고 75.2%의 간세포암종에서 관찰되었다. TGF-α 발현은 92.4%의 재생결절, 91.3%의 간세포종 그리고 93.2%의 간세포암종에서 관찰되었다. TGF-α의 발현을 나타내는 종양 중 68.0%에서 EGF-R의 발현을 나타내었다. ErbB2의 발현은 83.6%의 재생결절에서, 72.9%의 간세포종에서 그리고 88.7%의 간세포암종에서 관찰되었다. ErbB4 발현은 95.2%의 재생결절에서 86.3%의 간세포종에서 그리고 62.5%의 간세포암종에서 관찰되었으며, 간세포암종은 부분적인 발현 및 현저하게 감소된 발현 강도를 보였다. 결론: 본 연구의 결과에 따르면, EGF-R의 발현 증가와 ErbB4의 발현감소는 DEN에 의하여 유도된 간암 발생 모델에서 종양의 진전 과정과 관련이 있을 것으로 생각된다. 또한 인체 간질환과 유사한 실험 동물 모델 구축을 통하여 종양 표적 분자를 이용한 종양 치료 및 진전 예방의 방법 모색에 대한 기틀을 제공할 수 있다고 본다.
Abstract
Background/Aims: ErbB receptor proteins are transmembrane tyrosine kinase receptors; when they are activated by interaction with ligands, they generate diverse cellular responses, especially during lesion development and progression to cancer. In this study the expression of ErbB receptors and TGF-α were investigated using an experimental cirrhosis rat model giving rise to hepatocellular neoplasms, similar to human liver diseases. Methods: Fifty three male rats received intraperitoneal injection of diethylnitrosamine (DEN, 50 mg/kg), weekly for 18 weeks. Until the eighth week, two rats were sacrificed every two weeks and from the tenth to the eighteenth week, five rats were sacrificed weekly. Grossly, dyschromatic and dysmorphic nodules were counted and categorized into three groups: N1/N2/N3: 3 mm≤x<5 mm/5 mm≤x<10 mm/x≥10 mm in diameter. All nodules were examined, histologically. Antibodies for GSTp, TGF-α, EGF-R, ErbB2, ErbB3 and ErbB4 were used for immunohistochemistry. Results: The onset of cirrhoses was noted from the twelfth week. Preneoplastic foci, hepatocellular adenomas (HCA) and hepatocellular carcinomas (HCC) were noted from the second, eleventh and fifteenth week, respectively. The nodules (N1/N2/N3: 397/258/64) included regenerating nodule; RN (N1/N2/N3: 72.3%/15.9%/0%), HCA (N1/N2/N3: 27.2%/82.2%/7.6%) and HCC (N1/N2/N3: 0.5%/ 1.9%/92.4%). EGF-R was expressed in 12.5% of RN, 64.7% HCA and 75.2% HCC. TGF-α was expressed in 92.4% of RN, 91.3% HCA and 93.2% HCC. Sixty eight percent of TGF-α expressing nodules showed concurrent EGF-R expression. ErbB2 was expressed in 83.6% of RN, 72.9% HCA and 88.7% HCC. ErbB4 was expressed in 95.2% of RN, 86.3% HCA and 62.5% HCC. Conclusions: Increased expression of EGF-R and decreased expression of ErbB4, might be related with tumor progression during DEN-induced hepatocarcinogenesis
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