Evaluation of epithelial mesenchymal transition in patients with chronic obstructive pulmonary disease
© Sohal et al; licensee BioMed Central Ltd. 2011
Received: 8 August 2011
Accepted: 5 October 2011
Published: 5 October 2011
The reticular basement membrane (Rbm) in smokers and especially smokers with COPD is fragmented with "clefts" containing cells staining for the collagenase matrix-metalloproteinase-9 (MMP-9) and fibroblast protein, S100A4. These cells are also present in the basal epithelium. Such changes are likely hallmarks of epithelial mesenchymal transition (EMT). We aimed to confirm the epithelial origin of these Rbm cells, and to exclude potential confounding by infiltrating inflammatory cells.
Endobronchial biopsy sections from 17 COPD current smokers, with documented Rbm splitting and cellularity were stained for neutrophil elastase (neutrophil marker), CD68 (macrophage/mature fibroblasts), CD4+/CD8+ T lymphocytes, CD19 (B-cells), CD11c (dendritic cells/inflammatory cells), and S100 (Langerhans cells). The number of cells in the Rbm and epithelium staining for these "inflammatory" cell markers were then compared to numbers staining for S100A4, "a documented EMT epitope". Slides were double stained for S100A4 and cytokeratin(s).
In the basal epithelium significantly more cells stained for S100A4 compared to infiltrating macrophages, fibroblasts or immune cells: median, 26 (21.3 - 37.3) versus 0 (0 - 9.6) per mm, p < 0.003. Markedly more S100A4 staining cells were also observed in the Rbm compared to infiltrating macrophages, neutrophils, fibroblasts or immune cells or any sub-type: 58 (37.3 - 92.6) versus 0 (0 - 4.8) cells/mm Rbm, p < 0.003. Cells in the basal epithelium 26 (21.3 - 37.3) per mm) and Rbm (5.9 (2.3 - 13.8) per mm) frequently double stained for both cytokeratin and S100A4.
These data provide additional support for active EMT in COPD airways.
Keywordscytokeratin clefts epithelial mesenchymal transition (EMT) inflammatory cells and S100A4
One of the features of chronic inflammatory airway diseases, including COPD, is airway remodelling . In COPD, remodelling may occur as a response to smoking-induced damage to the airways, [2–4] but the details of structural changes and underlying mechanism are poorly described or understood. One potential mechanism contributing to airway fibrosis is transition of airway epithelial cells to a mesenchymal phenotype with myofibroblast characteristics, which then migrate into the lamina propria, a process termed epithelial mesenchymal transition (EMT) [5, 6]. There is little knowledge regarding the potential for EMT to occur in COPD airways.
In a recently published study,  we provided preliminary data suggesting that EMT is indeed likely in smokers' airways, but is especially active in the airways in current smoker - COPD subjects. These findings were based on 'classic' cell marker staining as used in other publications regarding EMT [8–12]. As a likely hallmark of EMT, we showed that the reticular basement membrane (Rbm) in smokers and COPD is highly fragmented, with elongated spaces or cracks, which we termed "clefts" [7, 13], again typical of EMT as described in the literature [10–12, 14, 15]. These elongated spaces and cracks or clefts in the Rbm were often not empty, but frequently contained cells, which we found were positive for the mesenchymal markers S100A4, vimentin and MMP-9. MMP-9 is a type IV collagenase, which increases in abundance during the process of EMT. Functionally MMP-9 is thought to assist epithelial cell migration by disrupting underlying basement membrane and Rbm and likely to be responsible for the Rbm fragmentation and cleft formation.
Although these changes which we have described are consistent with the presence of active EMT, we need further confirmation; in particular that the Rbm cells we have noted as expressing mesenchymal markers also express epithelial markers. It is also possible that the cell data could be confounded by us staining inflammatory cells infiltrating from below, which could be those positive for S100A4 and MMP-9.
In order to provide further evidence of a potential epithelial origin of cells in the Rbm of smoking COPD subjects, we have undertaken double immunostaining with both an epithelial and mesenchymal marker (cytokeratin-(s) and S100A4, respectively). S100A4 may also be expressed at least weakly by some inflammatory cells [16, 17]. We therefore, immunostained biopsies from smoker COPD patients for neutrophils, macrophages, fibroblasts, CD4+ and CD8+ lymphocytes, B cells and also dendritic cells and Langerhans cells.
Since our previous observations had shown that the putative changes of EMT were most marked in actively smoking COPD subjects, this confirmatory "proof-of-concept" study was limited to this most relevant clinical phenotypic group. We have previously studied normal controls and smokers with normal lung function as comparator groups , and the latter group also show similar Rbm changes but to a lesser extent., but there seemed little reason to include them in this follow up. Where we focus on where the signal is greatest.
In this study stored biopsies were selected from 6 of the seventeen COPD current smokers (COPD-CS) previously recruited using GOLD criteria ; all had signed informed consent for ongoing analysis of their biopsies (approved by the Human Research Ethics Committee Tasmania Network). Subjects with a history of other respiratory disorders were excluded. No subjects were receiving corticosteroids (oral or inhaled). These 6 chosen were not exception in any way from the COPD group as a whole, but all had the typical COPD biopsy appearance that we have interpreted as likely airway mucosal EMT.
Endobronchial biopsies were taken from near sub-segmental carinae of the right lower lobe (using alligator forceps, FB-15C; Olympus, Tokyo) .
Paraffin embedded sections of 3 μm and 50 μm apart mounted on each slide. At room temperature (RT) sections were stained with the following antibodies: anti-CD11c monoclonal (dendritic cells/inflammatory cells) (Abcam, cat no. ab52632, at 1:20,000 for 2 hours at RT), anti-CD4/CD8 monoclonal antibodies (Novocastra, cat no. NCL-CD4-IF6/cat no. NCL-CD8-4B11, at 1:15 for 2 hours), anti-CD68 monoclonal (macrophages and mature fibroblasts) (Dako, cat no. M0814, at 1:500 for 30 minutes), anti-CD19 monoclonal (B-cells) (Abcam, cat no. Ab31947, at 1:8000 for 2 hours), anti-neutrophil elastase monoclonal (Dako, cat no. Ab31947, at 1:500 for 1 hour), anti-S100 polyclonal (Langerhans cells) (Dako, cat no. Z0311 at 1:3000 for 1 hour) and anti-S100A4 polyclonal (Dako, cat no. A5114, 1:2500 for 90 minutes). Antibodies were visualised using horseradish peroxidase (HRP) conjugated DAKO Envision plus reagent (cat no. K4001, anti-mouse or K4003 anti-rabbit) and diaminobenzidine (DAB) for colour resolution (brown).
Double-staining for cytokeratin and S100A4 was performed using an anti-pan-cytokeratin monoclonal antibody (Dako, cat no. M3515 at 1:50 for 1 hour) and then secondarily bound using Dako "Real" (cat no. K5005) alkaline phosphatase reagent and visualised using Dako (cat no. K0640) liquid permanent red (mayer's haematoxylin was added at this point to elaborate nuclei) this was followed by the anti S100A4 polyclonal antibody (1:2000 for 90 minutes) that was visualised using HRP conjugated DAKO Envision plus (cat no. K4003) reagent and vina green for colour resolution (10 minutes at RT, Biocare, cat no. VG807 H, L). For the negative controls, matched sequential sections were stained with primary antibody replaced with a species-appropriate IgG1 at equivalent dilutions and conditions. We have published using these methods [7, 13, 19, 20].
Biopsy analysis and quantitation
Computer-assisted image analysis was performed (Leica DM 2500 microscope, Microsystems, Germany), Spot insight 12 digital camera and Image Pro V5.1 (Media Cybernetics, USA) software. For this analysis, we used representative sections (n = 6) from 17 different COPD current smokers for each of the markers of interest. All slides were counted in a single batch by a single experienced observer (SS) with quality assurance on randomly selected slides provided by a professional academic pathologist (HK).
After all counting had been completed, the data were analysed. As data were not normally distributed, results are presented as medians and ranges for cells in the basal epithelium and Rbm stained for each marker respectively, and analysed relative to S100A4 cell staining. Comparisons between different markers were undertaken using the Mann Whitney U test. Associations between variables were assessed using Spearman's rank test. Statistical analyses were performed using SPSS 15.0 for Windows, 2003, with a two-tailed P-value ≤ 0.05 being considered significant.
Demographic details of the subjects who participated in the current study (n = 6), compared to the full original population of patient volunteers from whom they were selected (n = 17)
(n = 17)
(n = 6)
GOLD I/GOLD II
Smoking pack years (Range)
Confirming epithelial origin of cells stained for S100A4 in the Rbm
Quantification of S100A4/cytokeratin stained cells and S100A4/cytokeratin double-stained cells in the basal epithelium and Rbm; data expressed as medians and ranges
No. of basal epithelial cells stained per mm of Rbm
No. of cells staining in Rbm per mm of Rbm
26.6 (21.3 - 37.3)
58.1 (37.3 - 92.6)
191.7 (135.3 - 297.6)*
4.3 (0.3 - 11.6)*
Cytokeratin-(s) & S100A4 double staining
26.6 (21.3 - 37.3) (basal epithelial cells +ve for S100A4 as shown above were all +ve for Cytokeratin-(s)
5.9 (2.3 - 13.8)
Comparative analysis of S100A4 staining with that for immune and inflammatory cells
Quantification of inflammatory cell markers and S100A4-stained cells in the basal epithelium and Rbm; data expressed as medians and ranges
No. of basal epithelial cells stained per mm of Rbm
No. of cells staining in Rbm per mm of Rbm
26.6 (21.3 - 37.3)
58.1 (37.3 - 92.6)
0 (0 - 5.8)*
0 (0 - 4.8)*
5.7 (0.9 - 9.6)*
0.3 (0 - 3.8)*
0 (0 - 0)*
0 (0 - 0)*
1 (0 - 1.8)*
0 (0 - 0.2)*
0.3 (0 - 2.1)*
0 (0 - 0.6)*
0 (0 - 0)*
0 (0 - 0)*
0 (0 - 1.1)*
0.1 (0 - 2.7)*
We have previously published that EMT could be an active process in human smokers' airways,  based on the demonstration of Rbm fragmentation and cleft formation and the presence of cells in both Rbm and basal epithelium staining for established markers of EMT [12, 15] such as S100A4 and MMP-9. These changes were present in airway biopsies from asymptomatic smokers with normal lung function, but were most pronounced in currently smoking COPD patients. This confirmatory study, although with limited numbers, focussed on this most relevant clinical phenotypic group, but is likely to be generalizable also to smokers with normal lung function.
Although we have provided rare, human data in human COPD, suggesting the potential for EMT,  this conclusion has been open to reviewer scepticism that the markers of EMT that we used, although regarded as 'classic' in the literature, [12, 15] were not qualified with further markers, especially showing that the Rbm cells express epithelial markers as well as mesenchymal markers. It is also been suggested that the Rbm cell data could be confounded by infiltration from below by inflammatory cells, which could at least potentially be positive for S100A4 and MMP-9. Our COPD subjects in this study were on as needed "relief" bronchodilator medication only, and it is unlikely that the discussed appearances are due to confounding by therapy, which we have deliberately tried to avoid as much as possible.
In the present study we have followed up our previous findings by demonstrating that cells in the Rbm positive for mesenchymal markers did frequently stain with an epithelial marker. Further, the present study illustrates that there are cells in the Rbm that double stain for cytokeratin-(s) and the "EMT marker" S100A4, representing further evidence of EMT in smoking-related COPD. Furthermore, our staining of tissue comprehensively for markers of a variety of inflammatory cells indicates that our data were not confounded by significant infiltration of the Rbm by inflammatory or immune cells.
Although only a relatively small number of cells in the Rbm could be demonstrated to stain with cytokeratin, it is possible that more (indeed most) of these Rbm cells are actually of epithelial origin, as it is well documented that during development of EMT, epithelial cells lose epithelial markers as they gain mesenchymal markers. Therefore, a cell which has gone through a complete transition will not express epithelial markers. It is, however, reassuring from the point of view of our hypothesis about active EMT in the airways in smokers and COPD, that we have been able to demonstrate at least a reasonable number of double-stained cells in the Rbm. Similarly, the frequent presence of cells in the basal epithelium staining for S100A4 is further evidence of EMT in these COPD airway biopsies. The data are consistent with a model where migratory cells gain the S100A4 mesenchymal marker early, but then rapidly loose epithelial markers as they migrate through the Rbm.
We found that there were very few CD8+ and CD4+ T lymphocytes and no CD19 B-cells in the Rbm and basal epithelium. Cells positive for CD19 were mainly in the lamina propria and hardly detectable in the basal epithelium or Rbm. We also confirmed that our Rbm cellular S100A4 staining was not confounded by infiltrating dendritic cells or Langerhans cells for which sections were stained for CD11c (dendritic cell/inflammatory cell marker)  and S100 (human Langerhans cell marker) . Although they were certainly present, there were relatively few such cells in the basal epithelium and Rbm compared to S100A4-stained cells. Most of the cells positive for the inflammatory cells studied were observed in the lamina propria below the Rbm, and not in the Rbm itself. EMT is a process defined by loss of epithelial markers and gain of mesenchymal markers. A function of this is that there is no single definitive marker for EMT. S100A4 is widely published as representing a reasonable marker for EMT [8–12]. We have also shown that these S100A4 positive cells in the Rbm also double-stain for another mesenchymal protein, vimentin,  widely used as an EMT marker [12, 15].
EMT has only recently been recognised in the human lung or airway, [6, 8] and now we provide further confirmatory evidence for its active presence in COPD airways . However, EMT is well described in lung embryogenesis,  metastatic malignant disease  and as part of the repair process in renal disease following tissue injury . Active EMT is indicated by the degradation of underlying epithelial basement membrane and transition of epithelial cells into mesenchymal cells with migratory potential, such that they move away from the epithelium in which they originated into deeper tissue [12, 15].
Active airway EMT in COPD could potentially relate to subsequent fibrotic activity in the sub-epithelial tissue, or just reflect severity of COPD pathology. COPD is a complex disease with physiologically a mixture of a variable emphysema component and intrinsic airway narrowing. Pathologically the airways are always abnormal and it is this component we have sampled from large airway biopsies. Airway obstruction is thought to be mainly in the small airways, but the pathology affects the whole bronchial tree [2–5]. There are in fact very few data available on fibrosis or alterations in fibroblast/fibrocyte populations in the airway in COPD. Our data taken together suggest that remodelling changes can be more exaggerated in established COPD and especially in current smokers [28, 29]. This area of pathology needs more specific study and remodelling changes need relating to changes in airway calibre and stiffness. Even if there are no immediate physiological consequences, there may well be implication for other COPD-associated processes and especially airway malignancy where the concurrence of EMT may be a significant part of tumour pathogenesis.
EMT has been described in metastatic malignant disease especially when associated with S100A4 expression. Epithelial cell nuclear expression of S100A4 strongly correlates with both active EMT and metastatic disease in the oncology literature [8, 30, 31]. Increased expressions of S100A4 and MMP-9 are observed in human non-small cell lung cancer (NSCLC) and have significant correlations with clinical and biological behaviour of such cancer cells [8, 30, 31]. Epidermal growth factor receptor (EGFR) is also over-expressed in many types of cancers, including NSCLC and we have shown that this is co-expressed in the epithelium as part of the whole 'COPD phenotype signal' . It has been shown that clinical responsiveness to EGFR inhibition using the "biologic" erlotinib is directly linked to the degree of EMT in NSCLC . Our findings of smoking-related active EMT may help understanding of why lung cancer is so common in smokers, but especially so in those with COPD where we have found EMT to be most marked.
Our study is of course limited by the fact that it was carried out in living human subjects and is based on 'static' pictures. Further study may require COPD model systems in animals and/or human cell culture. This might allow direct, vital imaging to observe dynamic real-time changes in cell phenotype and cellular migration during EMT.
It has been argued that Rbm fragmentation in smokers and COPD biopsies in our studies represents an "artefact" due to tissue processing. Against this is the fact that clefts contained cells, and also, to some extent vessels . Further, tissues from normal healthy controls  and asthmatics (data not shown) were processed in exactly the same way, but rarely showed any such Rbm fragmentation.
In summary, double staining of cells by both epithelial and mesenchymal markers in both the basal layer of the epithelium and within the fragmented Rbm is supportive of our hypothesis that EMT is indeed likely to be an active process in the airways in smokers, and especially those of current-smoking COPD patients. We also provided evidence that these S100A4 stained cells are not being confused with infiltrating inflammatory or immune cells. We conclude that the most likely origin of the S100A4 and cytokeratin dual positive cells apparent within the Rbm in COPD is from the airway epithelium, with this representing additional evidence that EMT may occur.
National Health and Medical Research Council (NHMRC) grant 490023 and Royal Hobart Hospital Research Foundation (RHHRF)
- Nakano Y, Muller NL, King GG, Niimi A, Kalloger SE, Mishima M, Pare PD: Quantitative Assessment of Airway Remodeling Using High-Resolution CT. Chest 2002,122(90060):271S-a-275.View ArticlePubMedGoogle Scholar
- Churg A, Tai H, Coulthard T, Wang R, Wright JL: Cigarette Smoke Drives Small Airway Remodeling by Induction of Growth Factors in the Airway Wall. Am J Respir Crit Care Med 2006,174(12):1327–1334.View ArticlePubMedGoogle Scholar
- Hogg JC, Macklem PT, Thurlbeck WM: Site and nature of airway obstruction in chronic obstructive lung disease. N Engl J Med 1968,278(25):1355–1360.View ArticlePubMedGoogle Scholar
- Saetta M, Turato G, Baraldo S, Zanin A, Braccioni F, Mapp CE, Maestrelli P, Cavallesco G, Papi A, Fabbri LM: Goblet cell hyperplasia and epithelial inflammation in peripheral airways of smokers with both symptoms of chronic bronchitis and chronic airflow limitation. Am J Respir Crit Care Med 2000,161(3 Pt 1):1016–1021.View ArticlePubMedGoogle Scholar
- Jeffery PK: Remodeling and Inflammation of Bronchi in Asthma and Chronic Obstructive Pulmonary Disease. Proc Am Thorac Soc 2004,1(3):176–183.View ArticlePubMedGoogle Scholar
- Willis BC, duBois RM, Borok Z: Epithelial Origin of Myofibroblasts during Fibrosis in the Lung. Proc Am Thorac Soc 2006,3(4):377–382.View ArticlePubMedPubMed CentralGoogle Scholar
- Sohal SS, Reid D, Soltani A, Ward C, Weston S, Muller HK, Wood-Baker R, Walters EH: Reticular basement membrane fragmentation and potential epithelial mesenchymal transition is exaggerated in the airways of smokers with chronic obstructive pulmonary disease. Respirology 2010,15(6):930–938.View ArticlePubMedGoogle Scholar
- Ward C, Forrest IA, Murphy DM, Johnson GE, Robertson H, Cawston TE, Fisher AJ, Dark JH, Lordan JL, Kirby JA, et al.: Phenotype of airway epithelial cells suggests epithelial to mesenchymal cell transition in clinically stable lung transplant recipients. Thorax 2005,60(10):865–871.View ArticlePubMedPubMed CentralGoogle Scholar
- Hodge S, Holmes M, Banerjee B, Musk M, Kicic A, Waterer G, Reynolds PN, Hodge G, Chambers DC: Posttransplant bronchiolitis obliterans syndrome is associated with bronchial epithelial to mesenchymal transition. Am J Transplant 2009,9(4):727–733.View ArticlePubMedGoogle Scholar
- Kalluri R: EMT: when epithelial cells decide to become mesenchymal-like cells. J Clin Invest 2009,119(6):1417–1419.View ArticlePubMedPubMed CentralGoogle Scholar
- Kalluri R, Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest 2003,112(12):1776–1784.View ArticlePubMedPubMed CentralGoogle Scholar
- Kalluri R, Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest 2009,119(6):1420–1428.View ArticlePubMedPubMed CentralGoogle Scholar
- Soltani A, Reid DW, Sohal SS, Wood-Baker R, Weston S, Muller HK, Walters EH: Basement membrane and vascular remodelling in smokers and chronic obstructive pulmonary disease: a cross-sectional study. Respir Res 2010,11(1):105..View ArticlePubMedPubMed CentralGoogle Scholar
- Acloque H, Adams MS, Fishwick K, Bronner-Fraser M, Nieto MA: Epithelial-mesenchymal transitions: the importance of changing cell state in development and disease. J Clin Invest 2009,119(6):1438–1449.View ArticlePubMedPubMed CentralGoogle Scholar
- Zeisberg M, Neilson EG: Biomarkers for epithelial-mesenchymal transitions. J Clin Invest 2009,119(6):1429–1437.View ArticlePubMedPubMed CentralGoogle Scholar
- Boomershine CS, Chamberlain A, Kendall P, Afshar-Sharif AR, Huang H, Washington MK, Lawson WE, Thomas JW, Blackwell TS, Bhowmick NA: Autoimmune pancreatitis results from loss of TGFbeta signalling in S100A4-positive dendritic cells. Gut 2009,58(9):1267–1274.View ArticlePubMedPubMed CentralGoogle Scholar
- Mazzucchelli L: Protein S100A4: too long overlooked by pathologists? Am J Pathol 2002,160(1):7–13.View ArticlePubMedPubMed CentralGoogle Scholar
- Rabe KF, Hurd S, Anzueto A, Barnes PJ, Buist SA, Calverley P, Fukuchi Y, Jenkins C, Rodriguez-Roisin R, van Weel C, et al.: Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary. Am J Respir Crit Care Med 2007,176(6):532–555.View ArticlePubMedGoogle Scholar
- Feltis BN, Wignarajah D, Zheng L, Ward C, Reid D, Harding R, Walters EH: Increased vascular endothelial growth factor and receptors: relationship to angiogenesis in asthma. Am J Respir Crit Care Med 2006,173(11):1201–1207.View ArticlePubMedGoogle Scholar
- Khor YH, Teoh AK, Lam SM, Mo DC, Weston S, Reid DW, Walters EH: Increased vascular permeability precedes cellular inflammation as asthma control deteriorates. Clin Exp Allergy 2009,39(11):1659–1667.View ArticlePubMedGoogle Scholar
- Beranek JT: CD68 is not a macrophage-specific antigen. Ann Rheum Dis 2005,64(2):342–343. author reply 343–344PubMedPubMed CentralGoogle Scholar
- Inoue T, Plieth D, Venkov CD, Xu C, Neilson EG: Antibodies against macrophages that overlap in specificity with fibroblasts. Kidney Int 2005,67(6):2488–2493.View ArticlePubMedGoogle Scholar
- Takano K, Kojima T, Go M, Murata M, Ichimiya S, Himi T, Sawada N: HLA-DR-and CD11c-positive dendritic cells penetrate beyond well-developed epithelial tight junctions in human nasal mucosa of allergic rhinitis. J Histochem Cytochem 2005,53(5):611–619.View ArticlePubMedGoogle Scholar
- Chon S, Kyung SY, Lee SP, Park JW, Jeong SH, Choi SJ, Ha SY: A case of pulmonary langerhans' cell histiocytosis mimicking hematogenous pulmonary metastases. Korean J Intern Med 2009,24(4):393–396.View ArticlePubMedPubMed CentralGoogle Scholar
- Lee JM, Dedhar S, Kalluri R, Thompson EW: The epithelial-mesenchymal transition: new insights in signaling, development, and disease. J Cell Biol 2006,172(7):973–981.View ArticlePubMedPubMed CentralGoogle Scholar
- Bjornland K, Winberg JO, Odegaard OT, Hovig E, Loennechen T, Aasen AO, Fodstad O, Malandsmo GM: S100A4 Involvement in Metastasis: Deregulation of Matrix Metalloproteinases and Tissue Inhibitors of Matrix Metalloproteinases in Osteosarcoma Cells Transfected with an Anti-S100A4 Ribozyme. Cancer Res 1999,59(18):4702–4708.PubMedGoogle Scholar
- Yanez-Mo M, Lara-Pezzi E, Selgas R, Ramirez-Huesca M, Dominguez-Jimenez C, Jimenez-Heffernan JA, Aguilera A, Sanchez-Tomero JA, Bajo MA, Alvarez V, et al.: Peritoneal Dialysis and Epithelial-to-Mesenchymal Transition of Mesothelial Cells. N Engl J Med 2003,348(5):403–413.View ArticlePubMedGoogle Scholar
- Dasari V, Gallup M, Lemjabbar H, Maltseva I, McNamara N: Epithelial-mesenchymal transition in lung cancer: is tobacco the "smoking gun"? Am J Respir Cell Mol Biol 2006,35(1):3–9.View ArticlePubMedGoogle Scholar
- Young RP, Hopkins R, Eaton TE: Pharmacological actions of statins: potential utility in COPD. Eur Respir Rev 2009,18(114):222–232.View ArticlePubMedGoogle Scholar
- Chen XL, Wang LC, Zhang WG, Chen XY, Sun ZM: [Correlations of S100A4 and MMP9 expressions to infiltration, metastasis and prognosis of non-small cell lung cancer]. Nan Fang Yi Ke Da Xue Xue Bao 2008,28(7):1254–1258.PubMedGoogle Scholar
- Flatmark K, Pedersen KB, Nesland JM, Rasmussen H, Aamodt G, Mikalsen SO, Bjornland K, Fodstad O, Maelandsmo GM: Nuclear localization of the metastasis-related protein S100A4 correlates with tumour stage in colorectal cancer. J Pathol 2003,200(5):589–595.View ArticlePubMedGoogle Scholar
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