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Lee, Han, Yang, Song, Yang, Kwon, Kang, Lee, Heo, Cho, and Song: The Incidence Rate and Severity of Orthotopic Lung Cancer in an Animal Model Depends on the Number of A549 Cells and Transplantation Period
Original Article

Lab. Anim. Res. 2010;26(4):369-375.

Published online: 20 January 2010

DOI: https://doi.org/10.5625/lar.2010.26.4.369

The Incidence Rate and Severity of Orthotopic Lung Cancer in an Animal Model Depends on the Number of A549 Cells and Transplantation Period

Jinsoo Lee1, 2, Young-Ah Han1, Hyo-Seon Yang1, Jeong-Ah Song1, Young-Su Yang1, Soonjin Kwon1, 2, Min-Sung Kang1, Kyuhong Lee1, Jeong-Doo Heo1, Kyu-Hyuk Cho1, Chang Woo Song1, ∗

1Inhalation Toxicology Center, Korea Institute of Toxicology Jeongeup Campus, Jeongeup, Korea

2Major of Pharmacology and Toxicology, University of Science and Technology, Daejeon, Korea

∗Corresponding author: Chang Woo Song, Inhalation Toxicology Center, Korea Institute of Toxicology, 1051 Shinjeong-dong, Jeongeup, Jeonbuk 580-185, Korea Tel: +82-63-570-8101 Fax: +82-63-570-8108 E-mail: cwsong@kitox.re.kr

Received 7 September 2010       Revised 23 November 2010       Accepted 29 November 2010

Copyright © 2010 Korean Association for Laboratory Animal Science

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The incidence rate of lung cancer is continually increasing, and lung cancer is the leading cause of cancer-related death worldwide. Nevertheless, few therapeutic methods are available for lung cancer. Therefore, establishing appropriate lung cancer animal models is important to investigate mechanisms and to evaluate new drugs for lung cancer. In the present study, we transplanted non-small cell lung cancer A549 human adenocarcinoma cells (2×104, 2.0×105, and 2.0×106 cells) into the right lobe of BALB/ c nude mice via the intercostal space to develop an orthotopic lung cancer animal model that is minimally invasive and similar to human lung cancer. We then investigated the incidence rate and severity of lung cancer according to the A549 cell number (2×104, 2.0×105, and 2.0×106 cells) and transplantation periods (4∼23 days). Lung cancer development was confirmed with gross examination, which was supported by histopathological examination. These results indicate that the incidence rate and severity of lung cancer was increased depending on the number of transplanted cells and transplantation period which the cell number and duration are increasing risk of lung cancer. Thus, this study can provide appropriate reference data to develop an orthotopic lung cancer animal model using the nonsmall cell lung cancer A549 cell line for researching mechanisms and evaluating candidate drugs, including various approaches for treating lung cancer.

Keywords: Lung cancer animal model, orthotopic, A549

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Figure 1.
Study design of an orthotopic lung cancer mouse model. (A) A549 adenocarcinoma cells (2.0×104) were transplanted into BALB/c nude mice. (B) A549 adenocarcinoma cells (2.0×105 and 2.0×106) were transplanted into BALB/c nude mice.
lar-26-369f1.tif
Figure 2.
Histopathological observation of the lung tissue. (A) Saline was injected into BALB/c nude mice at day 22. (B) A549 adenocarcinoma cells (2.0×106) were transplanted into BALB/c nude mice at day 22.
lar-26-369f2.tif
Figure 3.
Gross examination after transplantation of A549 adenocarcinoma cells, indicates that the severity of lung cancer (arrow) depends on the number of transplanted cells. (A) Saline was injected into BALB/c nude mice at day 22. (B) A549 adenocarcinoma cells (2.0×104) were transplanted into BALB/c nude mice at day 23. (C) A549 adenocarcinoma cells (2.0×105) were transplanted into BALB/c nude mice at day 22. (D) A549 adenocarcinoma cells (2.0×106) were transplanted into BALB/c nude mice at day 22.
lar-26-369f3.tif
Figure 4.
Histopathological examination after transplantation of A549 adenocarcinoma cells indicates that the severity of lung cancer (star) depends on the number of transplanted cells. (A) Saline was injected into BALB/c nude mice at day 22. (B) A549 adenocarcinoma cells (2.0×104) were transplanted into BALB/c nude mice at day 23. (C) A549 adenocarcinoma cells (2.0×105) were transplanted into BALB/c nude mice at day 22. (D) A549 adenocarcinoma cells (2.0×106) were transplanted into BALB/c nude mice at day 22.
lar-26-369f4.tif
Figure 5.
Gross examination after transplantation of A549 adenocarcinoma cells indicates that the severity of lung cancer (arrow) depends on the number of transplantation days. (A) Saline was injected into BALB/c nude mice at day 22. (B) A549 adenocarcinoma cells (2.0×106) were transplanted into BALB/c nude mice at day 8. (C) A549 adenocarcinoma cells (2.0×106) were transplanted into BALB/c nude mice at day 15. (D) A549 adenocarcinoma cells (2.0×106) were transplanted into BALB/c nude mice at day 22.
lar-26-369f5.tif
Figure 6.
Tumor area in histopathological slides depends on the number of transplanted A549 adenocarcinoma cells and transplanted days (mean±SE). Tumor area depends on transplanted days was significantly different (P<0.05). n=5∼6.
lar-26-369f6.tif
Table 1.
The incidence rate of lung cancer observed with gross and histopathological examination depends on the number of A549 adenocarcinoma cells
Examination/Cell amount Gross Histopathological
Day Saline a)2.0×104 b)2.0×105 b)2.0×106 Saline a)2.0×104 b)2.0×105 b)2.0×106
Day 4 0/5 0/5 – – 0/5 4/5 – –
Day 8 0/5 2/5 3/6 4/6 0/5 5/5 6/6 6/6
Day 15 0/6 – 6/6 6/6 0/6 – 6/6 6/6
Day 16 0/5 3/5 – – 0/5 5/5 – –
Day 22 0/6 – 6/6 6/6 0/6 – 6/6 6/6
Day 23 0/6 5/6 – – 0/6 6/6 – –

a) Gross and histopathological observation was performed after necropsy at day 4, 8, 16, and 23.

b) Gross and histopathological observation was performed after necropsy at day 8, 15, and 22.

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