窖蛋白-1在肺癌中的表达及意义

目的 探讨窖蛋白-1（caveolin-1）在不同类型肺癌组织中的表达及其与微血管密度（MVD）和临床病理因素之间的关系。方法 对154例原发性肺癌、相应癌旁正常肺组织及36例淋巴结转移癌行caveolin-1免疫组织化学染色;对154例原发性肺癌行CD34免疫组织化学（SP法）染色并进行微血管密度计数;Western印迹法检测其中50例新鲜肺癌组织及其癌旁正常肺组织中caveolin-1的表达情况。结果 caveolin-1为膜／质表达蛋白,在正常支气管上皮细胞和肺泡上皮细胞中的阳性率为100％。在肺癌组织中的阳性率为59．1％（91／154）,低于癌旁正常肺组织,P＜0．01;Western印迹结果进一步证实caveolin-1在肺鳞癌、肺腺癌组织中的表达均显著低于癌旁正常肺组织,P＜0．01。caveolin-1在小细胞肺癌（SCLC）和非小细胞肺癌（NSCLC）中的阳性率分别为7．1％和64．3％,二者间差异有统计学意义,P＜0．01。NSCLC中,有淋巴结转移组caveolin-1表达高于无淋巴结转移组（P=0．005）;Ⅲ、Ⅳ期组caveolin-1表达显著高于Ⅰ、Ⅱ期组（P=0．042）,caveolin-1表达与NSCLC的其他临床病理因素及MVD值无关（P＞0．05）。结论caveolin-1其作为一种肿瘤抑制因子的同时,可能还具有促进NSCLC进展和转移的活性。     

Localization of the 92 kd gelatinase mRNA in squamous cell and adenocarcinomas of the lung using in situ hybridization.

We have used in situ hybridization for RNA to localize cells containing mRNA for the 92 kd gelatinase in carcinomas of the lung. We used archival material to analyze sections from 12 cases of squamous cell carcinomas of the lung including six stage I and three stage II and from three cases of adenocarcinoma of the lung. Presence of mRNA in the tissue was verified by in situ hybridization for gamma actin. The 92 kd gelatinase mRNA was found in all 12 squamous cell carcinomas tumors and was highly expressed in the tumor cells themselves. In addition, it was found in host stromal cells surrounding the tumor, but not in normal lung fibroblasts. In contrast it was not found in the adenocarcinomas of the lung or in the stroma surrounding these tumors. The mRNA for the 92 kd gelatinase was present in normal pulmonary tissue, bronchial epithelium, basal cell hyperplasia of bronchial epithelium, alveolar macrophages, and focally in bronchial mucous glands. It was not present in normal alveoli, vascular cells, cartilage, or most lymphocytes. We corroborated the presence of the mRNA for the 92 kd gelatinase by ribonuclease protection assay. The levels of mRNA for the 92 kd gelatinase in two specimens of squamous cell carcinoma were 6- to 10-fold greater than in the nonneoplastic tissue and two adenocarcinoma specimens.     

基因芯片检测人肺鳞癌和肺腺癌基因表达的异同

目的用基因芯片技术检测肺鳞癌和肺腺癌基因表达的异同。方法提取人肺鳞癌和肺腺癌组织及正常肺组织的RNA,分别用Cy5-dCTP或Cy3-dCTP标记,再与4096点基因芯片杂交,检测肺鳞癌和肺腺癌组织基因表达的异同。结果肺鳞癌和肺腺癌表达共同上调的基因17条,共同下调的基因19条;肺鳞癌表达显著高于肺腺癌的基因20条,显著低于肺腺癌的基因14条。结论多基因参与肺癌发病,基因芯片技术是肺癌基因表达检测的有效方法。     

Retinoic acid receptor-beta expression in stage I non-small cell lung cancer and adjacent normal appearing bronchial epithelium

Retinoic acid receptor-beta (RAR-beta) is induced by and mediates the growth-inhibitory and apoptotic effects of retinoic acid (RA), suggesting that loss of RAR-beta expression may be one of the critical events involved in the carcinogenesis/ progression of non-small cell lung cancer (NSCLC) and in the responsiveness to retinoid chemotherapy. However, recent contradictory reports that the expression of RAR-beta is associated with poor clinical outcome, and the fact that treatment of serum-deprived type 2 alveolar cells with RA leads to a stimulation of cell proliferation, require the verification of RAR-beta as a biomarker of chemoprevention or prognosis. The expression status of RAR-beta in cancer cells and adjacent normal appearing bronchial epithelium from 39 patients, diagnosed as stage I NSCLC and undergone a curative lung resection, was analyzed in paraffin-embedded tissue sections by IHC staining. The normal appearing bronchial epithelium of 14 out of 33 (42.4%) specimens expressed RAR-beta, whereas 22 out of the 39 (56.4%) stage I NSCLC specimens expressed RAR-beta. RAR-beta was more frequently expressed in the adenocarcinoma (72.7%) than in the squamous cell carcinoma (31.3%) (p=0.026). Neither the expression status in normal appearing adjacent tissue nor that in the tumor tissue had prognostic implications. The higher expression of RAR-beta in cancer tissue, the focal and uneven distribution in normal appearing adjacent bronchial epithelium, and inconsistency with the corresponding tumor tissue, suggest that the expression status of RAR-beta as a biomarker for chemoprevention/early diagnosis or the prognosis of NSCLC requires further consideration.     

ARHGAP10基因在肺腺癌中的表达及其机制

目的:基于生物信息学技术探讨Rho GTPase活化蛋白10（ARHGAP10）在肺腺癌发生发展中的潜在分子机制,为后续深入研究提供生物信息学证据。方法:通过利用TCGA数据库,获取肺腺癌患者RNA高通量序列及相关患者的临床资料,其中包括514例肺腺癌患者的癌组织以及59例相关的癌旁组织;GSE115002肺腺癌基因芯片数据从GEO下载,芯片包含52例肺腺癌患者的癌组织和匹配的癌旁正常组织。通过R语言技术比较ARHGAP10在TCGA和GSE115002中肺腺癌组织和癌旁正常组织之间表达差异,并进一步分析ARHGAP10表达与肺癌患者临床预后的相关性;挖掘两个数据库中ARHGAP10重叠基因,对重叠基因进行GO和KEGG富集分析,采用STRING数据库获取ARHGAP10的相关基因互作蛋白网络,TIMER数据库分析ARHGAP10与免疫浸润的相关性。结果:与癌旁正常组织相比较,肺腺癌组织中ARHGAP10呈现低表达状态（P<0.01）。与ARHGAP10低表达组相比较,ARHGAP10高表达患者拥有更长的总生存期和无进展生存期（P<0.05）。TCGA和GSE115002肺腺癌数据库中ARHGAP10的共同相关基因共133个,与MAPK、PI3K-AKT等信号通路以及细胞外基质受体结合、肌动蛋白细胞骨架、肿瘤信号通路等生物学进程相关。本研究发现PRELP、LIMS2、PPAP2B、MRPL42、SRP9、TSNAX等蛋白在ARHGAP10相互作用网络中发挥重要作用,ARHGAP10与DC细胞、中性粒细胞浸润正相关,与PD-L1表达呈正相关。结论:ARHGAP10在肺腺癌中起抑癌作用,与肺腺癌的生存预后正相关,可作为肺腺癌患者潜在的临床预后标志物。     

Relation of fine structure to prognosis for papillary adenocarcinoma of the lung

Eight cases of papillary adenocarcinoma of the lung were investigated by light and electron microscopy. Prognoses for all but one patient were favorable. Two patients (husband and wife) who underwent tumor resection during the same year experienced no disease-related problems for more than seven years afterward. Microscopic study of the papillary adenocarcinomas revealed columnar, peg-shaped, mucus-secreting tumor cells lining the alveoli. The tumor tissue contained alveolar macrophages, with some multinucleated giant cells, interstitial lymphoid infiltrates, mildly thickened alveolar walls, and a few pneumoconiotic foci. These inflammatory stromal reactions in the tumor tissue might have been associated with the favorable prognoses. Nuclear inclusions were detected in some tumor cells in all cases. Ultrastructurally, the inclusions contained tubular, granular, crystalline, and electron-dense homogeneous structures in addition to unclassifiable nuclear bodies. The tubular structures were in close proximity to inner leaflets of the nuclear membranes. The papillary adenocarcinoma cells had features of totipotential bronchioloalveolar cells, differentiating toward type II pneumocytes, Clara cells, and ciliated epithelial cells.     

Expression profile of early lung adenocarcinoma: identification of MRP3 as a molecular marker for early progression

Early lung adenocarcinoma is well-recognized as a small-sized non-invasive adenocarcinoma or localized non-mucinous bronchioloalveolar carcinoma (LNMBAC); however, the molecular events associated with these early lesions are not clear. To determine the genes involved in tumorigenesis at the early stage of lung adenocarcinoma, we compared the mRNA expression profiles of LNMBAC and normal lungs with an oligonucleotide array. Immunohistochemical analyses were performed to confirm the expression of detected genes. We identified 183 differentially expressed genes, of which 15 were up-regulated and 168 down-regulated. Among them, most up-regulated genes, such as AQP3 and Claudin-4, were expressed in both adenocarcinoma cells and type II alveolar pneumocytes, corresponding to the histological similarity between these cell types. However, multidrug resistant protein 3 (MRP3) was only expressed on tumour cell membranes and not in type II alveolar pneumocytes, as confirmed by immunohistochemistry. Moreover, the number of MRP3-positive cells significantly increased from AAH (the precursor lesion of lung adenocarcinoma) to LNMBAC. We conclude that MRP3 could be a novel molecular marker for LNMBAC, whose expression increases during the early progression of tumourigenesis.     

hnRNP B1 expression in benign and malignant lung disease

Evidence is accumulating to suggest that hnRNP B1 expression may be a useful tool in the early diagnosis of lung cancer. This study examined the immunohistochemical expression of hnRNP B1 in archived sections of resected lung cancers and compared the patterns of expression with those seen in similar archived sections of non-neoplastic lung. Particular attention was paid to the expression of hnRNP B1 in the benign bronchial cells in both cases, to establish if overexpression of this protein in respiratory epithelial cells is specific for malignancy. Nineteen cases of different types of non-small cell carcinoma were examined (eight squamous cell, six adenocarcinomas, two carcinosarcomas, two undifferentiated large cell carcinomas, and one mucoepidermoid carcinoma) and compared with sections from 16 open lung biopsies (three cases of cryptogenic fibrosing alveolitis, two cases of sarcoidosis, two cases of organizing pneumonia, and one case each of tuberculosis, extrinsic allergic alveolitis, non-specific interstitial pneumonitis, pneumocystis pneumonia, aspergilloma, respiratory bronchiolitis-interstitial lung disease, mineral dust disease, Sj?gren's syndrome and systemic sclerosis vascular variant). All the tumours showed positive staining, with the vast majority, 16/19 (84%), showing strong diffuse nuclear staining. The background cells of these cases showed positive staining in alveolar macrophages, lymph node germinal centres, bronchial mucous glands, and bronchial epithelial cells. No significant difference was seen in the percentage of positive bronchial epithelial cells in bronchi adjacent to the tumour compared with the resection margins. In the benign lung cases, positive bronchial epithelial cells were seen in a small percentage, 3/16 (18%), of cases, but the majority of cases showed no or very focal staining. The levels of expression between benign epithelial cells of malignant cases, compared with benign, showed a significant difference when the staining was assessed in percentage of positive nuclei (p = 0.001, Fisher's exact test). The results confirm that hnRNP B1 is widely expressed in a range of lung carcinomas; that expression is seen in benign bronchial epithelial cells and inflammatory cells; and that expression in background bronchial epithelial cells appears to be higher in malignant than in benign lung disease. It is feasible that this biomarker may be of use in the detection of early lung cancer, provided that levels of expression can be accurately quantified.     

肺腺癌中miRNA34a和c-myc的表达及意义

目的检测肺腺癌组织中miRNA34a,癌基因c-myc的表达,探讨两者之间的关系及意义。方法采用qRT-PCR法检测mRNA miRNA34a,Western blot、免疫组化法检测c-myc蛋白在正常肺组织、癌旁肺组织、肺腺癌以及转移癌组织中的表达。结果(1)miRNA34a在肺腺癌组织中呈低表达,低于癌旁和正常肺组织(P0.01),高于转移癌组织(P0.01)。c-myc在肺腺癌组织中呈高表达,高于癌旁和正常肺组织(P0.01),低于转移癌组织(P0.01)。miRNA34a及c-myc在癌旁组织和正常肺组织之间的表达差异有统计学意义(P0.05)。(2)病理类型:miRNA34a及c-myc在低分化和高分化肺腺癌之间的表达差异均无统计学意义(P0.05)。两者在转移癌与非转移肺腺癌组织中的表达差异有统计学意义(P0.05)。(3)预后分析:cmyc阳性组的肺腺癌患者生存率低于阴性组(P0.05),miRNA34a低表达组患者的生存时间明显低于高表达组,差异有统计学意义(P0.01)。结论肺腺癌组织中miRNA34a的表达与c-myc蛋白表达呈负相关。c-myc高表达miRNA34a低表达提示患者预后不良。     

Decreased Beclin-1 expression is correlated with the growth of the primary tumor in patients with squamous cell carcinoma and adenocarcinoma of the lung

Beclin-1 is a Bcl-2-interacting protein, and it may delay cell cycle progression and induce autophagy. The function and expression of Beclin-1 and Bcl-2 in squamous cell carcinoma and adenocarcinoma of the lung remain largely unknown. Herein, we investigated the expression of Beclin-1 and Bcl-2 in squamous cell carcinoma and adenocarcinoma of the lung. Tissue samples from 262 cases were used in this study. Immunohistochemical staining for Beclin-1 and Bcl-2 were conducted using a tissue microarray. In squamous cell carcinoma, Beclin-1 expression was strongly positive in 48 (28.6%) of 168 samples, it was moderately positive in 42 (25.0%) of 168 samples, and it was negative or weakly positive in 78 (46.4%) of 168 samples. In adenocarcinoma, Beclin-1 expression was strongly positive in 26 (27.7%) of 94 samples, it was moderately positive in 27 (28.7%) of 94 samples, and it was negative or weakly positive in 41 (43.6%) of 94 samples. Beclin-1 expression was inversely correlated with the tumor size and the primary tumor stage (pT) in both types of tumor. Especially, the TNM stage of adenocarcinoma was inversely correlated with Beclin-1 expression. Our results suggest that a progressively reduced Beclin-1 expression is correlated with the primary tumor growth of squamous cell carcinoma and adenocarcinoma of the lung.     

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     

Clinicopathology of stromal invasion in lung adenocarcinoma

In the World Health Organization classification, lung adenocarcinoma with mixed subtypes is defined as invasive carcinoma with evidence of vascular, pleural, or stromal invasion. The histological criteria for stromal invasion, however, are not clearly established. A total of 157 peripheral pure bronchioloalveolar carcinoma (BAC) or lung adenocarcinoma with mixed BAC and others were reviewed. All cases had been resected between 1986 and 2000 and measured ≤30 mm in maximum dimension. Destruction of alveolar framework (DAF) was defined as distortion or discontinuity of the alveolar framework by tumor growth. The extra-alveolar area involvement (EAAI) was defined as tumor growth outside the alveolar framework, which includes the following areas: bronchial wall, perivascular connective tissue and/or the vascular wall, interlobular septum and the visceral pleura. Survival of patients with adenocarcinoma without DAF ( n  = 41) was 100%. Even when adenocarcinoma involved DAF and lacked EAAI ( n  = 21), survival was 100%. The 5 year survival rate of groups with two invasion signs ( n  = 34) was 90.1%, and that of groups with three to five invasion signs ( n  = 61) was 66.7%. Tumor growth outside the alveolar framework is the hallmark of stromal invasion.     

OV-TL 12/30 (keratin 7 antibody) is a marker of glandular differentiation in lung cancer

The immunoreactivity of OV-TL 12/30, a monoclonal antibody to keratin 7 was investigated on paraffin-embedded human lung cancer tissues of 61 patients. A modified AEC-immunoperoxidase method with pepsin pre-digestion was used. In normal lung tissue keratin 7 was found in bronchial and bronchiolar epithelium, pneumocytes and compound glands. Squamous metaplasia of the bronchial tree was negative. All 24 squamous cell carcinomas were negative irrespective of grade of differentiation. All differentiation grades of 20 adenocarcinomas including bronchioalveolar carcinomas were positive. Since six large cell anaplastic carcinomas did not react with keratin 7 antibody these tumours are considered to be of squamous cell rather than adenocarcinomatous origin. Small cell anaplastic carcinomas were negative in 10 of 11 cases. Our study demonstrates that this keratin 7 antibody is useful in differentiating between squamous cell carcinoma and adenocarcinoma of the lung and it may be particularly useful in making the correct diagnosis in small lung biopsy specimens.     

MADD在肺腺癌组织中的表达及对肺腺癌A549细胞增殖和凋亡的影响

目的:探讨MADD在肺正常组织及肺腺癌组织中的表达及其对肺腺癌细胞增殖和凋亡的影响。方法:收集肺临床病理组织标本,免疫组化法检测肺正常组织和肺癌组织MADD表达;培养人肺腺癌A549细胞,逆转录PCR检测其IG20基因表达;用携带MADD基因的质粒和能够沉默MADD表达的慢病毒载体分别转染A549细胞,Western blot、MTT分析和流式细胞术检测其MADD表达、增殖及凋亡。结果:肺腺癌组织MADD表达水平明显高于肺正常组织和肺鳞癌组织;A549细胞能够表达MADD;高表达MADD能抑制A549细胞凋亡,提高其增殖活力,而沉默MADD表达则能促进A549细胞凋亡,降低其增殖活力。结论:肺腺癌组织MADD表达明显增高;MADD可通过抑制凋亡来促进肺腺癌细胞生存。     

Therapeutic effect of anti CEACAM6 monoclonal antibody against lung adenocarcinoma by enhancing anoikis sensitivity

Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) plays a crucial role in tumorigenesis of lung cancer. However, the therapeutic potential for anti CEACAM6 monoclonal antibody (mAb) has only been limitedly explored. Here, we evaluate the therapeutic potential of naked anti CEACAM6 mAb against lung adenocarcinoma. Clone 8F5, recognizing B domain of CEACAM6, is established by immunizing A549 cells and screening for clones double positive for A549 and CEACAM6-Fc recombinant protein. We found that 85.7% of 70 resected lung adenocarcinoma tissue sections were positive for CEACAM6, whereas all squamous cell carcinoma examined were negative. A549 cells with high levels of CEACAM6 demonstrated more aggressive growth nature and showed increased paclitaxel chemosensitivity upon 8F5 binding. Treatment with 8F5 to A549 decreased cellular CEACAM6 expression and reversed anoikis resistance. 8F5 also decreased cellular status of Akt phosphorylation and increased apoptosis via caspase activation. In a mouse model of lung adenocarcinoma with xenotransplanted A549 cells, 8F5 treatment alone demonstrated 40% tumor growth inhibition. When combined with paclitaxel treatment, 8F5 markedly enhanced tumor growth inhibition, up to 80%. In summary, we demonstrate that anti CEACAM6 mAb is an effective therapeutic treatment for lung adenocarcinoma whose effect is further enhanced by combined treatment with paclitaxel.     

Involucrin in lung tumors. A specific marker for squamous differentiation

Involucrin is a precursor of the cross-linked envelope protein or marginal band present in human stratum corneum. This study uses immunohistochemical techniques for localization of involucrin in histologic sections from 91 lung tumors in order to evaluate the usefulness of involucrin as a tumor marker in lung neoplasms. Although involucrin is absent from bronchial epithelium, it is expressed in cultured tracheal epithelial cell colonies and in bronchial mucosa with squamous metaplasia. Involucrin was present in all 25 cases of squamous and adenosquamous carcinoma. Staining was focal in 12 cases of squamous cell carcinoma and was most marked in the larger neoplastic cells in the center of squamous cell nests. Only two of 20 cases of adenocarcinoma revealed focal staining for involucrin, and these cases may represent adenosquamous variants. Six of 12 cases of large cell undifferentiated carcinoma stained for involucrin, indicating squamous differentiation, and seven cases of malignant mesothelioma were negative. Isolated involucrin-positive cells were present in two of 16 cases of small cell anaplastic carcinoma and one of 11 carcinoid tumors, identifying variants of neuroendocrine tumors with dual differentiation. Patterns of localization of involucrin in paraffin and frozen sections were compared with staining for cytokeratins in parallel sections. Immunohistochemical localization of involucrin comprises a specific marker for squamous differentiation in lung tumors.     

Wip1在肺癌细胞中的作用机制初探

目的: 检测Wip1及相关蛋白在肺癌细胞中的表达情况,探讨Wip1在肺癌发生中的作用机制。方法: Western blotting检测各种细胞(肺腺癌细胞A549、肺鳞癌细胞NCI-1299、大细胞肺癌细胞 NCI-H460和正常支气管上皮细胞HBE)中Wip1、 p53、p-p53、p38 MAPK和p-p38 MAPK蛋白的表达。结果: A549、NCI-1299和H460细胞中Wip1蛋白的表达均高于HBE细胞(P<0.05);各种肺癌细胞间Wip1蛋白的表达无明显差异(P>0.05);p-p53和p-p38 MAPK蛋白在肺癌细胞中的表达低于HBE细胞(P<0.05);肺癌细胞中Wip1表达增加,p-p38 MAPK和p-p53表达降低,存在着Wip1-p38 MAPK-p53信号通路的失衡。结论: 肺癌细胞(A549、NCI-1299、H460)中存在Wip1-p38 MAPK-p53信号通路的失衡。这可能是Wip1在肺癌中的致癌机制之一。