Protein 1 and Clara cell 10-kDa protein distribution in normal and neoplastic tissues with emphasis on the respiratory system

Thirty-six different normal tissues and 13 different malignant epithelial tumours, were examined immunohistochemically for the presence of protein 1 (P1) and Clara cell 10-kDa protein (CC10). Adenocarcinomas of the lung were also examined for the expression of pulmonary surfactant apoprotein using a monoclonal antibody (PE-10). The staining results of P1 and CC10 were almost identical both in normal tissues and in malignant tumours. In normal lung, Clara cells were strongly positive for both P1 and CC10. In addition, some goblet cells and non-ciliated non-mucus cells in the upper airways were moderately positive for both proteins. In the malignant tumours, some lung cancers were positive for P1 and CC10, both of which were positive in the same tumour cells on sequential sections. In 117 lung cancers, P1 and CC10 were positive in 10.2% of adenocarcinomas, 20.5% of squamous cell carcinomas, and 12.5% of large cell carcinomas. PE-10 stained positively in 65.3% of adenocarcinomas, a frequency significantly higher than that of P1 and CC10 (P<0.01). These results suggest that P1 and CC10 are nearly identical proteins, that both are useful markers of Clara cells, and that many pulmonary adenocarcinomas express surfactant apoprotein rather than Clara cell proteins.     

A case of pulmonary sclerosing hemangioma

A case of pulmonary sclerosing hemangioma was studied using light microscopic, immunohistochemical and electron microscopic methods. The cytoplasm of the sclerosing hemangioma cells was positive to the anti-lung surfactant apoprotein monoclonal antibody (PE-10). These cells included pale cells of the solid areas, cells covering papillary projections, and cells lining cleft-like spaces. Electron microscopic study showed that the predominant cells included poorly-differentiated pneumocytes and normal Type 2 pneumocytes. We concluded that the sclerosing hemangioma was an epithelial tumor with differentiation towards Type 2 pneumocytes. A part of this study was presented at the 25th Annual Meeting of the Clinical Electron Microscopy Society of Japan, Matsumoto, September 28–30, 1993.     

The expression of osteopontin and its association with Clara cell 10 kDa protein in allergic rhinitis

Background Osteopontin (OPN) is a multifunctional protein that has recently been linked to allergic diseases. Clara cell 10 kDa protein (CC10) is another protein linked to allergy, and has been suggested to have an inhibitory role in inflammatory airway diseases. At this time, it is not known whether OPN is involved in allergic rhinitis (AR) or if there is any association between CC10 and OPN in AR. Objective To study the expression of OPN and its potential association with CC10 in AR. Methods The expression of CC10 and OPN in nasal mucosa of AR patients was investigated. AR animal models were established by using wild‐type and CC10‐knockout mice. In some experiments, human recombinant CC10 protein was given to AR mice during either sensitization or challenge. The phenotypic changes were examined by histology and real‐time RT‐PCR. The direct effect of CC10 on the OPN expression in spleen mononuclear cells and on the OPN‐induced inflammatory cytokine expression in BEAS‐2B cells was measured through in vitro cell culture. Results OPN expression was up‐regulated, with a concomitant down‐regulation of CC10, in AR patients, showing a significant negative correlation between their expression. Compared with control mice sensitized with PBS, the OPN expression was significantly increased in AR mice; such an increase was more prominent in CC10‐knockout mice, compared with wild‐type. Administration of CC10 during both sensitization and challenge could markedly ameliorate Th2‐skewed inflammation and OPN expression in nasal mucosa. CC10 administration at the sensitization phase could also reduce spleen OPN expression. The in vitro study showed that CC10 directly down‐regulated the OPN expression in spleen mononuclear cells stimulated with OVA and suppressed the OPN‐induced expression of Th2 cytokines and pro‐inflammatory cytokines in BEAS‐2B cells. Conclusion In the context of allergic airway responses, CC10 can inhibit OPN expression and suppress the Th2‐promoting function of OPN, resulting in CC10's inhibitory biological effects. Cite this as: Y. Liu, X. Lu, H.‐J. Yu, X.‐Y. Hua, Y.‐H. Cui, S.‐K. Huang and Z. Liu, Clinical & Experimental Allergy, 2010 (40) 1632–1641.     

Combined small cell carcinoma with pulmonary blastoma and adenocarcinoma: case report and clonality analysis

We describe a rare tumor occurring in the left pulmonary lobe of a 71-year-old Japanese man. The tumor, which was resected by left lower lobectomy, measured 65×50×50 mm. Histologic examination revealed papillary adenocarcinoma in small cell carcinoma, and chondrosarcoma. Also, the blastemal cells were located between the small cell carcinoma and the chondrosarcoma, and intermingled with both components. In blastemal cells, some glands resembled a well-differentiated fetal adenocarcinoma. The tumor was diagnosed as combined small cell carcinoma with pulmonary blastoma and papillary adenocarcinoma according to the 2004 WHO classification. Immunohistochemically, the small cell carcinoma expressed TTF-1, pancytokeratin, CD56, synaptophysin, and S100 protein, while blastemal cells expressed vimentin, desmin, smooth muscle actin, CD56, and S100 protein. To investigate whether the tumor was clonal or not, p53 gene mutation of each tumor component was analyzed by laser-captured microdissection, polymerase chain reaction-single-strand conformation polymorphism and direct sequencing. Despite the histologic complexity, all components showed the same mutation at exon5 of the p53 gene. These results indicate that the tumor was clonal and arose from a relatively primitive cell, and that p53 mutation occurred before histologic metamorphosis or differentiation.     

Epithelial-mesenchymal transition in patients of pulmonary adenocarcinoma: correlation with cancer stem cell markers and prognosis

Adenocarcinoma is the most common histologic type of non-small cell lung carcinomas. The existence of lung cancer stem cells (CSCs) and epithelial-mesenchymal transition (EMT) in human tissue is controversy. The aim of this study is to investigate the expression and clinical significance of CSCs and EMT markers and evaluate the correlation between the two in lung adenocarcinoma. A total of 97 cases comprise the tissue microarray from surgical resection for primary lung adenocarcinoma. Immunohistochemistry for ALDH1 and CD44 as CSC markers and E-cadherin, vimentin, fibronectin, SMA as EMT markers was performed. High ALDH1A1 expression was statistically associated with female gender (P=0.001), smoker (P=0.012), and high pT stages (P=0.046). High CD44 expression was statistically associated with female gender (P=0.008), non-smoker (P=0.000), and no pleural invasion (P=0.039). High expression of ALDH1 was associated with good overall survival (P=0.021). High expression of CD44 was correlated with both good overall survival (P=0.024) and disease-free survival (P=0.000). Vimentin expression was associated with pT stage (P=0.001) and pleural invasion (P=0.028). E-cadherin, fibronectin and SMA were not associated with clinicopathologic correlation and all EMT markers were not correlated with survival of lung adenocarcinoma. CSC markers expression was not related to EMT. Our results showed that the expression of CSCs was associated with a good prognosis in lung adenocarcinoma. The prognostic significance of EMT markers was skeptical in this study. There is a need for more research about CSC, EMT, and the relation between these two in human lung adenocarcinoma.     

Inhaled LPS induces blood release of Clara cell specific protein (CC16) in human beings

BACKGROUND: Animal models of lung inflammation have validated the plasma 16-kd Clara cell protein (CC16) as a peripheral marker of the permeability of the alveolocapillary barrier. OBJECTIVE: We investigated in human beings whether inhaled LPS induced a rise in airways permeability measured by the plasma changes in CC16. METHODS: The CC16 was measured in plasma from 15 subjects exposed to LPS by inhalation, during which the kinetics and the dose-response relationship of LPS-induced CC16 were evaluated. Because LPS-induced response involves macrophages activation, the protective effect of oral methylprednisolone was also evaluated. RESULTS: An inhalation of 50 microg LPS induced a significant ( P < .001) rise in CC16 after 6 hours (from 7.24 [+/-0.68] microg/L to 10.69 [+/-0.99] microg/L) that normalized at 24 hours (6.65 [+/-0.33] microg/L). The CC16 response was dose-related, with the no-response threshold 0.5 microg LPS. A 6-day treatment with 20 mg/d methylprednisolone inhibited significantly ( P < .001) the CC16 response to 50 microg LPS. CONCLUSION: Exposure to LPS by inhalation in healthy subjects induces an intravascular leakage of CC16 that can be blocked by corticosteroids. These observations further validate plasma CC16 as a noninvasive test of the alveolocapillary barrier permeability.     

HMGB2对肺腺癌细胞周期和增殖的影响

目的:探讨高迁移率族蛋白B2(HMGB2)对肺腺癌细胞周期和增殖的影响。方法:从Cancer RNA-Seq Nexus (CRN)数据库分析肺腺癌组织HMGB2表达情况;从OncoLnc数据库分析HMGB2与肺腺癌患者预后的相关性;从肿瘤单细胞数据库(CancerSEA)分析HMGB2与肺腺癌细胞14种功能状态的相关性;利用siRNA技术下调人肺腺癌A549细胞中HMGB2表达,通过real-time PCR和Western blot验证沉默效果,CCK8和EdU实验检测细胞的增殖。结果:HMGB2在肺腺癌中高表达;HMGB2高表达组肺腺癌患者的总生存期明显低于HMGB2低表达患者(log-rank检验P=0.017 3);HMGB2表达与肺腺癌细胞周期和增殖呈正相关;敲减HMGB2表达后A549细胞的活力和增殖能力显著降低(P<0.05)。结论:HMGB2的表达与肺腺癌细胞周期和增殖显著正相关,可以作为潜在的评估肺腺癌患者预后的标志物和治疗靶标。     

Pulmonary epithelial expression of human α1-antitrypsin in transgenic mice results in delivery of α1-antitrypsin protein to the interstitium

α₁-Antitrypsin (α₁AT) therapy is used as a treatment for α₁AT deficiency. It has also been proposed as a therapy for cigarette smoke-induced emphysema, although the efficacy of such therapy is as yet unproven. Moreover, the optimal route of delivery of α₁AT to the lung interstitium, the crucial locus of action, is unknown. We created transgenic mice with expression of the human α₁AT gene directed by a human surfactant protein C (SpC) promoter fragment or a rat Clara cell 10-kDa protein (CC10) promoter fragment in order to examine the ability of pulmonary epithelial cell expression of α₁AT to deliver protein to the interstitium, and to produce a model that would allow studies on the efficacy of α₁AT in preventing lung damage after cigarette smoke exposure. Four transgenic lines were studied. In situ hybridization and light microscopic immunohistochemistry showed that two CC10 driven lines expressed human α₁AT in type II alveolar cells and airway epithelial cells; α₁AT expression was seen in the alveolar parenchyma in two SpC driven lines, and in small airway epithelium in one of the SpC lines. Electron microscopic immunochemistry showed the presence of the human α₁AT protein in the interstitium in all lines. Mean levels of human protein varied from 0.37 to 2.9 μg/g lung protein and serum levels from 0.72 to 1.3 μg/ml, compared to normal human serum α₁AT levels of 2–5 mg/ml. We conclude that transgene-mediated expression of α₁AT in pulmonary epithelial cells results in diffuse expression of the transgene in the alveolar parenchyma and reproducibly leads to transfer of protein to the interstitium. The present model is, however, limited by low levels of protein production; limited protein production may be a problem in other forms of gene therapy in which relatively large amounts of extracellular protein are needed in the lung for a therapeutic effect. Received: 5 August 1998 / Accepted: 25 January 1999     

Clear cell adenocarcinoma with endobronchial polypoid growth

Clear cell adenocarcinoma of the lung is extremely rare. On radiography, a 45-year-old female with fever was found to have an abnormal shadow in the left lower lung field. Bronchoscopy revealed a polypoid tumor in the left bronchus. On biopsy, the tumor was determined to be adenocarcinoma. Preoperative examination found no tumors outside of the lung. The patient underwent left lower lobectomy with bronchial wedge resection. The tumor had completely obstructed and dilated the left lower bronchus, but had not invaded the tissue outside the bronchial wall. Microscopically, the cytoplasm of the tumor cells contained abundant glycogen, and the tumor had solid and glandular structures. The tumor was diagnosed as clear cell adenocarcinoma of the lung.     

Clara cell secretory protein (CC16): characteristics and perspectives as lung peripheral biomarker

Clara cell protein (CC16) is a 15.8-kDa homodimeric protein secreted in large amounts in airways by the non-ciliated bronchiolar Clara cells. This protein increasingly appears to protect the respiratory tract against oxidative stress and inflammation. In vitro, CC16 has been shown to modulate the production and/or the activity of various mediators of the inflammatory response including PLA2, interferon-gamma and tumour necrosis factor-alpha. CC16 has also been found to inhibit fibroblast migration or to bind various endogenous or exogenous substances such as polychlorobiphenyls (PCBs). This protective role is confirmed by studies on transgenic mice, showing that CC16 deficiency is associated with an increased susceptibility of the lung to viral infections and oxidative stress. In humans, a polymorphism of the CC16 gene, localized to a region linked to airway diseases, has recently been discovered in association with an increased risk of developing childhood asthma. Finally, CC16 also presents a major interest as a peripheral marker for assessing the integrity of the lung epithelium. The determination of CC16 in serum is a new non-invasive test to detect Clara cell damage or an increased epithelial permeability in various acute and chronic lung disorders.     

Association of small foci of diffuse idiopathic pulmonary neuroendocrine cell hyperplasia (DIPNECH) with adenocarcinoma of the lung

DIPNECH is regarded as a precursor lesion of neuroendocrine lung tumors, specifically carcinoids. A relationship with lung adenocarcinomas has not been clearly established so far. We present a series of four cases with a concomitant presence of adenocarcinoma and DIPNECH in the lung.     

Ultrastructure of Well-Differentiated Adenocarcinomas of the Lung with Special Reference to Bronchioloalveolar Carcinoma

The cytologic phenotypes of 20 well-differentiated pulmonary adenocarcinomas were determined by electron microscopy. On examination of more than 100 cells in each case, the tumors were classified according to the predominant cell types. Nine cases (45%) were of mucous cell type, further divided into 7 cases of bronchial surface epithelial cell type, 1 case of bronchial gland cell type, and 1 case of metaplastic bronchiolar goblet cell type. The remainder included 5 cases (25%) of Clara cell type, 2 cases (10%) of type II cell type, and 4 cases (20%) of mixed cell type. The predominant histologic pattern by light microscopy was “typically” bronchioloalveolar (Manning et al.'s type 1) in the metaplastic goblet cell tumor and papillary in most Clara cell-type tumors, while it was glandular in bronchial surface and bronchial gland cell types, although variable in type II cell or mixed cell type. Therefore, bronchioloalveolar carcinomas, when histologically defined inclusive of papillary tumors, present cytologic phenotypes also related to the bronchioloalveolar epithelium, i.e., metaplastic goblet or Clara or type II cell subtypes, which is in accordance with some previous reports. These tumors could be distinguished from the other (glandular) adenocarcinomas that show primarily bronchial mucous cell differentiation.     

Frequent appearance of club cell (Clara cell)‐like cells as a histological marker for ALK‐positive lung adenocarcinoma

Anaplastic lymphoma kinase‐rearranged (ALK⁺) lung cancers show characteristic histological features, such as solid signet ring cell patterns and mucinous cribriform patterns; however, these features are not always observed in ALK⁺ lung cancers. We noticed that club cell (Clara cell)‐like cells (CLCs) were frequently present in the papillary portion of ALK⁺ lung adenocarcinomas. In this study, we investigated the importance of CLCs in papillary patterns of ALK⁺ lung cancers. We compared the histological features of 18 ALK⁺ cases with 62 control cases (22 epidermal growth factor receptor‐positive (EGFR⁺) and 40 ALK‐ and EGFR‐negative (ALK^?/EGFR^?) cases). The present study analyzed presence of papillary pattern, proportion of papillary pattern area, presence of micropapillary pattern, frequency of CLCs and lengths of snout. The frequency of CLCs in ALK⁺ cases was significantly higher than that in EGFR⁺ cases and ALK^?/EGFR^? cases. Micropapillary pattern was more frequently observed in ALK⁺ cases than that in ALK^?/EGFR^? cases (P < 0.001). The present study indicated that the high frequency of CLCs in papillary patterns was significantly associated with ALK⁺ cases. When solid signet ring cell patterns and mucinous cribriform patterns are absent, the high frequency of CLCs in papillary adenocarcinoma could be a useful histological marker for ALK⁺ lung cancers.     

Expression patterns of type II pneumocyte apical surface glycoconjugates in lung adenocarcinoma cells

 Monoclonal antibodies and lectins were used to examine the expression patterns of apical membrane oligosaccharide sequences specific to type II pneumocytes in atypical adenomatous hyperplasia (AAH) and lung cancer. Atypical cells of AAH and papillary adenocarcinoma cells expressed abundant sialyl Thomsen-Friedenreich (TF) antigen: this was not observed in acinar adenocarcinoma, bronchioloalveolar carcinoma with mucin production or squamous cell carcinoma. Sialyl Tn antigens was also detected on a few cells in AAH and papillary adenocarcinomas. Asialo TF and Tn antigen were not observed on the surface of carcinoma cells of any type. Alpha(α)2,3-linked sialic acids predominated in type II pneumocyte, AAH and papillary adenocarcinoma, whereas ciliated columnar cells expressed α2,6-linked sialic acids. Lewis^x and sialyl Lewis^x antigens capped the TF antigen in both O- and N-linked side chains on the surface of AAH and papillary adenocarcinoma cells, but were not expressed by type II pneumocytes. The findings demonstrate that papillary adenocarcinoma cells resemble type II pneumocytes in that they express abundant sialyl TF surface antigen, but they also express TF-related antigens not found in type II pneumocytes. Apical surface glycoconjugates of AAH have structural characteristics shared by both type II pneumocytes and papillary adenocarcinoma cells. Received: 6 July 1998 / Accepted: 25 September 1998     

Pulmonary adenocarcinoma with enteric differentiation: Dissecting oncogenic genes alterations with DNA sequencing and FISH analysis

Background

Pulmonary Adenocarcinoma with Enteric Differentiation (PAED) is a rare subtype of adenocarcinoma of emerging interest, recently introduced in the 2015 WHO classification. However, little is known about major molecular signatures of this class of adenocarcinomas and information about new biomarkers totally lack.