Chest
Volume 135, Issue 2, February 2009, Pages 344-352
Journal home page for Chest

Original Research
COPD
Proteomic Analysis in Lung Tissue of Smokers and COPD Patients

https://doi.org/10.1378/chest.08-1583Get rights and content

Rationale

Although cigarette smoking is the most important risk factor for COPD, COPD develops in only a minority of smokers, suggesting a significant genetic role. To solve the underlying pathophysiologic mechanism, it is critical to understand genes and their final product, ie, proteins. We investigated the exclusive proteins from the lung tissues obtained from COPD patients using proteomics.

Methods

Nontumorous lung tissue specimens were obtained from patients who underwent surgery for lung cancer. We included 22 subjects: nonsmokers (n = 8), smokers without COPD (healthy smokers, n = 7), and smokers with COPD (n = 7). Proteins were separated from their spots with two-dimensional polyacrylamide gel electrophoresis and examined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). To validate the proteins from the above procedures, Western blotting and immunohistochemistry were conducted.

Results

Twelve protein spots from COPD group significantly increased or decreased compared with the other two groups were chosen for MALDI-TOF-MS analysis. Eight proteins were up-regulated in the COPD group as compared with the nonsmokers. Meanwhile, five proteins from the COPD group were up-regulated and five were down-regulated when compared with healthy smokers. Of these, matrix metalloproteinase (MMP)-13 and thioredoxin-like 2 were significantly increased in the COPD patients by Western blot and immunohistochemistry. MMP-13 was mainly expressed in the alveolar macrophages and type II pneumocytes; however, thioredoxin-like 2 was primarily seen in the bronchial epithelium.

Conclusions

MMP-13 and thioredoxin-like 2 in lungs increased in patients with COPD. MMP-13 was mainly expressed in the alveolar macrophages and type II pneumocytes. In contrast, thioredoxin-like 2 was primarily seen in the bronchial epithelium.

Section snippets

Study Subjects

Nontumorous lung tissue specimens were obtained from patients who underwent lobectomy or pneumonectomy for lung cancer. After surgical resection, these lung tissues were immediately frozen to − 80°C; they were then donated and stored at the Korea Lung Tissue Bank, which has been assigned and supported by the Korean Science and Engineering Foundation in the Ministry of Sciences and Technology. All patients included in the study gave their consent, and the study protocol was approved by the

Two-Dimensional Polyacrylamide Gel Electrophoresis

The COPD group revealed a mean age of 56.0 years (mean tobacco smoking history, 38.3 pack-years). Mean FEV1 was 66.1% of predicted in the COPD patient group. Similarly, mean age in the nonsmoker group was 59.0 years, and mean age and mean pack-years of the healthy smoker group were 64.5 years and 48.7 pack-years, respectively. Age, sex, smoking history, and pulmonary function data of patients are summarized in Table 1.

Figure 1A shows an example of the increased expression in COPD patients as

Discussion

In this study, we have shown that the expressions of MMP-13 and thioredoxin-like 2 were increased in the COPD lung tissues. MMP-13 was mainly expressed in the alveolar macrophages and type II pneumocytes. In contrast, thioredoxin-like 2 was primarily seen in the bronchial epithelium.

MMP-13, one of the proteolytic enzymes, is thought to be mainly expressed by chondrocytes and synovial cells in human osteoarthritis and rheumatoid arthritis; it has a role in cartilage destruction. MMP-13 also is

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    This work was supported by grant R21-2007-000-10058-0 from the National Research Resource Bank Program.

    This work was performed at the Division of Respiratory and Critical Care Medicine, Department of Internal Medicine, Korea University College of Medicine, Seoul, Korea.

    The authors have no conflicts of interest to disclose.

    Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (www.chestjournal.org/misc/reprints.shtml).

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