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Shin, Lee, and Park: Expression of Caspase 3, Survivin, and p53 Protein in Urethane Induced Mouse Lung Carcinogenesis
Original Article
Tuberculosis and Respiratory Diseases

Tuberculosis and Respiratory Diseases 2007; 63(3): 251-260.

Published online: 30 September 2007

DOI: https://doi.org/10.4046/trd.2007.63.3.251

Expression of Caspase 3, Survivin, and p53 Protein in Urethane Induced Mouse Lung Carcinogenesis

Jong Wook Shin, M.D.1, Soo Hwan Lee, M.D.2, Eon Sub Park, M.D.2

1Division of Allergy, Respiratory and Critical Care Medicine, Department of Internal Medicine, Chung Ang University College of Medicine, Seoul, Korea.

2Department of Pathology, Chung Ang University College of Medicine, Seoul, Korea.

Address for correspondence: Jong Wook Shin1, M.D. and Eon Sub Park2, M.D. 1Division of Allergy, Respiratory and Critical Care Medicine, Department of Internal Medicine, 2Department of Pathology, Chung-Ang University Hospital, 224-1, Huk-suk-dong, Donjak-gu, Seoul, Korea. Phone: 82-2-6299-1407, Fax: 82-2-6263-2184, basthma@cau.ac.kr, basthma@hanmail.net, esp@cau.ac.kr

Received 3 May 2007       Accepted 14 September 2007

Copyright © 2007 The Korean Academy of Tuberculosis and Respiratory Diseases

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.

Keywords: Survivin, caspase-3, p53, Lung cancer, Lung carcinogenesis, PCNA, Apoptosis

Figures and Tables

Figure 1
A. Gross finding of urethane-induced lungs, which shows multiple masses. B, C. Sequential change in urethane-induced lung lesions, showing adenoma (Figure 1B) and infiltrating adenocarcinoma (Figure 1C)(Hematoxylline & Eosin Stain, ×200). D. Immunohistochemical reactivity for PCNA in adenoma (×200). E. TUNNEL stain shows occasional nuclear staining in adenocarcinoma of lung (×200). F, G, H. Immunohistochemical reactivity for capase 3 (Figure 1F), p53 (Figure 1G) and survivin protein (Figure 1H) showing positive cytoplasmic staining (×200).
trd-63-251-g001
Table 1
Histologic changes of the lung in mice treated with urethane
trd-63-251-i001

Tumor multiplicity; Number of tumor per mouse.

Table 2
PCNA and apoptotic labeling index of the mice lung lesions during urethane-induced carcinogenesis
trd-63-251-i002

Significantly greater than the other groups, p<0.01.

Table 3
Expression rates of caspase-3, survivin and p53 protein in mouse lung lesions by urethane-induced lung carcinogenesis
trd-63-251-i003

NS: not significantly different.

Table 4
Proliferation and apoptotic index according to caspase 3 and survivin expressin expression
trd-63-251-i004

n: number of tumor lesion; NS: not significantly different. Significantly greater than the other groups, p<0.01.

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