bromochloroacetic-acid and Small-Cell-Lung-Carcinoma

Small cell lung carcinoma (SCLC), a malignancy of neuroendocrine origin, can show varied morphologies and patterns but is typically positive for at least one neuroendocrine marker and almost always for cytokeratins. It is essential to distinguish this tumour due to its characteristic genetic features, aggressive behaviour, propensity for metastasis and responsiveness to chemotherapy. We hereby present a rare case of a pulmonary mass that showed morphological features of an SCLC but lacked cytokeratin expression on biopsy as well as resection specimens. Various cytokeratins were tested on multiple blocks and at different laboratories. A broad differential diagnosis was considered and ruled out including small round blue cell tumours, non-SCLC and metastasis. After performing an extensive work-up to identify the origin of this tumour, it was finally diagnosed as SCLC with expression of neuroendocrine markers synaptophysin and CD56, and intracytoplasmic electron dense neurosecretory granules (250-350 nm) however lacked cytokeratin expression.

Topics: Biomarkers, Tumor; Biopsy; Carcinoma, Neuroendocrine; Carcinoma, Non-Small-Cell Lung; Humans; Keratins; Lung Neoplasms; Small Cell Lung Carcinoma

Fibroblast growth factor receptor 1 (FGFR1) is frequently amplified in human small-cell lung cancer (SCLC), but its contribution to SCLC and other lung tumors has remained elusive. Here, we assess the tumorigenic capacity of constitutive-active FGFR1 (FGFR1

Topics: Adenocarcinoma of Lung; Animals; Bronchi; Cell Transformation, Neoplastic; Cisplatin; Drug Resistance, Neoplasm; Gene Expression Regulation, Neoplastic; Humans; Integrases; Keratins; Lung Neoplasms; Mice; Mutation; Nasal Cavity; Neurosecretory Systems; Oncogenes; Pulmonary Alveoli; Receptor, Fibroblast Growth Factor, Type 1; Retinoblastoma Protein; Small Cell Lung Carcinoma; Tumor Suppressor Protein p53

Small cell lung carcinoma (SCLC) is an aggressive pulmonary neoplasm of neuroendocrine origin. Keratins form a large group of intermediate filaments, which are major structural proteins in epithelial cells and carcinomas. SCLC shows a wide spectrum of keratin expression, from very strong to completely negative. A prognostic role of keratin expression in SCLC is unknown.. Tumor tissue microarray samples from a unique series of 82 SCLC patients who underwent pulmonary resection were stained with keratin specific antibodies AE1/AE3 and CAM5.2. The percentage o1f positively stained cells and their staining pattern (diffusely membranous, partially membranous and dot-like) were evaluated. The median expression value was used for the distinction between keratin-negative and -positive patients. Overall survival in respective groups was compared using the log-rank test. Multivariate Cox proportional hazards regression analysis was performed adjusting for age, gender, tumor site, tumor stage, and tumor histology.. edian expression of AE1/AE3 and CAM5.2 was 80% and 90%, respectively. Five cases were completely negative for AE1/AE3 and three for Cam5.2. Median overall survival for patients with stronger and weaker AE1/AE3 staining was 24.7 and 13.8 months, respectively (p=0.019). There was no difference in survival in relation to the CAM5.2 expression (p=0.44). In multivariate analysis adjusted for CAM5.2, T and N stage, gender and age at diagnosis, stronger AE1/AE3 expression was an independent predictor of increased survival (HR 0.50; 95% CI, 0.27-0.94; p=0.031).. High expression of AE1/AE3 is a favorable prognostic factor in surgically treated SCLC. The applicability of this finding to a typical patient population treated with non-surgical methods warrants further studies.

Topics: Antiporters; Biomarkers; Female; Humans; Keratins; Lung; Male; Multivariate Analysis; Prognosis; Proportional Hazards Models; Small Cell Lung Carcinoma; Tissue Array Analysis

Small cell lung cancer (SCLC) is characterized by prevalent circulating tumour cells (CTCs), early metastasis and poor prognosis. We show that SCLC patients (37/38) have rare CTC subpopulations co-expressing vascular endothelial-cadherin (VE-cadherin) and cytokeratins consistent with vasculogenic mimicry (VM), a process whereby tumour cells form 'endothelial-like' vessels. Single-cell genomic analysis reveals characteristic SCLC genomic changes in both VE-cadherin-positive and -negative CTCs. Higher levels of VM are associated with worse overall survival in 41 limited-stage patients' biopsies (P<0.025). VM vessels are also observed in 9/10 CTC patient-derived explants (CDX), where molecular analysis of fractionated VE-cadherin-positive cells uncovered copy-number alterations and mutated TP53, confirming human tumour origin. VE-cadherin is required for VM in NCI-H446 SCLC xenografts, where VM decreases tumour latency and, despite increased cisplatin intra-tumour delivery, decreases cisplatin efficacy. The functional significance of VM in SCLC suggests VM regulation may provide new targets for therapeutic intervention.

Topics: Animals; Antigens, CD; Biopsy; Cadherins; Cell Line, Tumor; Cohort Studies; DNA Copy Number Variations; Female; Humans; Keratins; Lung; Lung Neoplasms; Male; Mice; Middle Aged; Mutation; Neoplastic Cells, Circulating; Neovascularization, Pathologic; Single-Cell Analysis; Small Cell Lung Carcinoma; Tumor Suppressor Protein p53; Xenograft Model Antitumor Assays

Patients with small cell lung carcinoma (SCLC) rarely demonstrate long-term survival. We previously reported that gene expression profiling identified a subset of SCLC with good prognosis in surgical cases. To find an easier way to routinely identify SCLC belonging to this subset, we conducted the present study with a hypothesis that neuroendocrine (NE) or basaloid (BA) phenotypes may influence prognosis. To confirm the subset, we used an array platform to analyze fresh samples. Because inoperable cases may differ from surgical cases, we enrolled 51 biopsy cases and 43 resected samples. To evaluate NE and BA phenotypes, we used NE (synaptophysin, chromogranin A, and CD56) and BA (p63 and CK34βE12) markers. To varying extents, expression profiling based on the array platform duplicated the subsets. For NE phenotypes, 77% of surgical cases and 100% of biopsy cases were positive for at least 1 marker. For BA phenotypes, only 19% of surgical cases were positive for at least 1 marker, whereas there were no positive biopsy cases. Cases undergoing surgery were categorized based on NE and BA immunoreactivity; 58% into NE+BA-, 19% into NE+BA+, 23% into NE-BA-, and 0 into NE-BA+ groups. NE- patients (n = 10) demonstrated a significantly better prognosis (P = .0306) than their NE+ counterparts (n = 33), whereas no survival difference was evident between the BA+ and BA- groups. Multivariate analyses showed that NE positivity was an independent prognostic factor. In conclusion, the SCLC subset with good prognosis is identified by low NE marker expression, which was found only in surgical cases.

Topics: Aged; Biomarkers, Tumor; Biopsy; Carcinoma, Neuroendocrine; CD56 Antigen; Chromogranin A; Female; Gene Expression Profiling; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Membrane Proteins; Middle Aged; Prognosis; Protein Array Analysis; Small Cell Lung Carcinoma; Synaptophysin

The use of p63 has been advocated for separating small cell lung carcinoma from poorly differentiated non-small cell lung carcinoma, in particular, squamous cell lung carcinoma. However, p63 is not entirely specific in this distinction, as several studies have demonstrated p63 immunoreactivity in a proportion of small cell lung carcinomas. p40 (ΔNp63) has recently been purported to be a highly specific marker for squamous cell lung carcinoma, but data regarding p40 (ΔNp63) in small cell lung carcinoma, a key differential diagnostic consideration in biopsy samples of squamous cell lung carcinoma, are lacking. In this study, a total of 34 previously confirmed small cell lung carcinomas (27 bronchoscopic biopsy samples and 7 large specimens) were immunostained for p40 (ΔNp63), p63, and keratin 34βE12. All 34 small cell lung carcinomas were negative for p40 (ΔNp63) and keratin 34βE12. Although none of the large small cell lung carcinoma specimens exhibited p63 immunoreactivity, 12 (44.4%) of 27 biopsy samples of small cell lung carcinoma were positive for p63. The rate of p63 staining in small cell lung carcinoma biopsy samples differed significantly from that of p40 (ΔNp63) and keratin 34βE12 (P = .005). Ten cases of squamous cell lung carcinoma were also tested, all of which were positive for all 3 markers. These findings confirm that p63 immunoexpression is not uncommon in biopsy samples of small cell lung carcinoma. Positive p63 staining may be mistakenly interpreted as supportive of squamous differentiation and result in misclassification of small cell lung carcinoma as squamous cell lung carcinoma, a diagnostic error that has important therapeutic and prognostic consequences. To provide greater diagnostic accuracy, p40 (ΔNp63) or keratin 34βE12 should be used instead of p63 in the distinction of small cell lung carcinoma from non-small cell lung carcinoma in biopsy samples.

Topics: Biomarkers, Tumor; Biopsy; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Small Cell Lung Carcinoma

Epithelial-to-mesenchymal transition (EMT), the phenotypical change of cells from an epithelial to a mesenchymal type, is thought to be a key event in invasion and metastasis of adenocarcinomas. These changes involve loss of keratin expression as well as loss of cell polarity and adhesion. We here aimed to determine whether the loss of keratin expression itself drives increased invasion and metastasis in adenocarcinomas and whether keratin loss leads to the phenotypic changes associated with EMT. Therefore, we employed a recently described murine model in which conditional deletion of the Keratin cluster II by Cre-recombinase leads to the loss of the entire keratinmultiprotein family. These mice were crossed into a newly generated Cre-recombinase inducible KRAS-driven murine lung cancer model to examine the effect of keratin loss on morphology, invasion and metastasis as well as expression of EMT related genes in the resulting tumors. We here clearly show that loss of a functional keratin cytoskeleton did not significantly alter tumor morphology or biology in terms of invasion, metastasis, proliferation or tumor burden and did not lead to induction of EMT. Further, tumor cells did not induce synchronously expression of vimentin, which is often seen in EMT, to compensate for keratin loss. In summary, our data suggest that changes in cell shape and migration that underlie EMT are dependent on changes in signaling pathways that cause secondary changes in keratin expression and organization. Thus, we conclude that loss of the keratin cytoskeleton per se is not sufficient to causally drive EMT in this tumor model.

Topics: Animals; Biomarkers; Carcinoma, Non-Small-Cell Lung; Cell Line; Disease Models, Animal; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Gene Order; Gene Targeting; Genes, ras; Humans; Keratins; Lung Neoplasms; Mice; Mice, Transgenic; Small Cell Lung Carcinoma

Topics: Aged; Biomarkers, Tumor; Diagnosis, Differential; Fatal Outcome; Humans; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Lymphatic Metastasis; Neoplasm Proteins; Small Cell Lung Carcinoma

Lung cancer cells express several cytokeratins (CKs) that are subdivided into type I (CK9-23) and type II (CK1-8) subclasses. The functions of CKs in lung cancer cells have not been fully elucidated. The purpose of this study was to investigate the role of CKs in the invasion of lung cancer cells. We investigated the expression levels of CK7, 8, 18 and 19 in 12 non-small cell lung cancer (NSCLC) and seven SCLC cell lines by quantitative immunoblotting. The expression levels of these four CKs were significantly higher in the NSCLC cells. The NSCLC cell line HI1017 expressed CK8 and 18; A549 cells expressed CK7, 8, 18 and 19, respectively. Invasive sublines of HI1017 and A549 were established by repeated selection of invasive cells using a membrane invasion chamber system. The invasive cell lines showed lower expression levels of CKs compared with the parental cells. Exogenous CK19 also resulted in a decrease in invasiveness of the HI1017 cells. Suppression of either CK8 or CK18 by short interfering RNAs led to a decrease in the total CKs and increased invasiveness of both the HI1017 and A549 cells. A549 cells expressed very low levels of CK19. Suppression of CK19 affected neither invasive ability nor total CK amount in the A549 cells. Our observations indicate that CK expression levels were inversely associated with invasiveness of the NSCLC cell lines, and suggest that expression levels of dominant CKs may affect invasive ability.

Topics: Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Humans; Immunoblotting; Keratins; Lung Neoplasms; Neoplasm Invasiveness; RNA Interference; Small Cell Lung Carcinoma

The aim of the present study was to clarify protein profiling in small cell lung carcinoma (SCLC) and pulmonary large cell neuroendocrine carcinoma (LCNEC). The proteomic approach was used, and involved cell lysate from two cell lines (N231 derived from SCLC and LCN1 derived from LCNEC), with 2-D gel electrophoresis (2-DE). In the present study, 25 protein spots with greater than twofold quantitative differences between LCN1 and N231 cells on 2-DE gels were confirmed. Within the 25 identified proteins, cytokeratins (CK) 7, 8, 18 and 19 were upregulated in LCN1 cells compared with N231 cells. The expression of CK7, 8, 18, and 19 was further studied on immunohistochemistry with 81 formalin-fixed and paraffin-embedded pulmonary carcinomas, which included 27 SCLC, 30 LCNEC, 14 adenocarcinomas, and 10 squamous cell carcinomas. Although the expression of CK7, 8, 18, and 19 was observed in all histological types, the mean immunostaining scores of CK7, 8, 18, and 19 were significantly higher in LCNEC than in SCLC (P < 0.001, P < 0.001, P < 0.01 and P < 0.001, respectively). These data suggest that the biological characteristics of LCNEC and SCLC may be different and the expression of CK may serve as differential diagnostic markers.

Topics: Biomarkers, Tumor; Carcinoma, Large Cell; Carcinoma, Neuroendocrine; Electrophoresis, Gel, Two-Dimensional; Humans; Immunohistochemistry; Keratin-18; Keratin-19; Keratin-7; Keratin-8; Keratins; Lung Neoplasms; Small Cell Lung Carcinoma; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tissue Array Analysis

To investigate the clinical significance of a combination of several serum tumor markers in the diagnosis of lung cancer.. Serum CEA, CA125, CA199, CYFRA21-1 and NSE were measured with RIA, chromatometrychemoluminescence, ELISA and biochemoluminescence methods respectively in 340 patients with lung cancers at different TNM stages, 120 patients with benign lung diseases, and 45 healthy people. The sensitivities, specificities and accuracies of different combination of those markers for the diagnosis of lung cancers were compared.. CYFRA21-1 had the highest sensitivity and accuracy (60.0% and 70.3%) and CA199 had the highest specificity (99.4%) for detecting lung cancers. CYFRA21-1 had the highest sensitivity (79.6%) for detecting squamous carcinoma. CEA had the highest sensitivity of (75.7%) for detecting adenocarcinoma. NSE had the highest sensitivity (76.2%) for detecting small cell lung cancers (SCLC). The combination of several serum tumor markers had higher sensitivities than the single marker for the diagnosis of lung cancers. With two markers, the combination of CYFRA21-1 and NSE had the highest sensitivity and accuracy (82.9% and 83.4%), while the combination of CA125 and CA199 had the highest specificity (94.5%). With three markers, the combination of CEA, NSE and CYFRA21-1 had the highest sensitivity and accuracy (89.1% and 85.1%), while the combination of CEA, CA125 and CA199 had the highest specificity (86.7%). The combination of CEA, CA125, CA199, CYFRA21-1 and NSE produced the best value, with a sensitivity of 93.8%, a specificity of 71.5%, and an accuracy of 86.5%.. Serum CEA, CA125, CA199, CYFRA21-1 and NSE are helpful markers for the diagnosis of lung cancer. The combination of the markers can improve the sensitivity and accuracy of the diagnosis.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; CA-125 Antigen; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Diagnosis, Differential; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Small Cell Lung Carcinoma

Besides new therapeutic drugs, effective diagnostic tools indicating early the efficacy of therapy are required to improve the individual management of patients with nonoperable cancer diseases.. In prospectively collected sera of 128 patients with newly diagnosed small cell lung cancer receiving first-line chemotherapy, the courses of nucleosomes, progastrin-releasing peptide (ProGRP), neuron-specific enolase (NSE), cytokeratin-19 fragments (CYFRA 21-1), and carcinoembryonic antigen were investigated and correlated with therapy response objectified by computed tomography before start of the third treatment course.. In univariate analyses, high levels and insufficient decreases of nucleosomes, ProGRP, NSE, and CYFRA 21-1 during the first and second cycles of therapy correlated with poor outcome. Insufficient response to therapy was most efficiently indicated by the baseline values of nucleosomes, ProGRP, and CYFRA 21-1 before the second therapy cycle reaching areas under the curve (AUC) of 81.8%, 71.3%, and 74.9% in receiver operating characteristic curves, respectively. Combinations of nucleosomes with ProGRP (AUC 84.1%), CYFRA 21-1 (AUC 82.5%), and NSE (AUC 83.6%) further improved the diagnostic power in the high specificity range and yielded sensitivities of 47.1%, 35.3%, and 35.3% at 95% specificity, respectively. In multivariate analyses, including clinical and biochemical variables, only performance score and nucleosomes before cycle 2 were found to independently indicate therapy response.. Biochemical markers specifically identified patients with insufficient therapy response at the early treatment phase and showed to be valuable for diseases management of small cell lung cancer.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Antineoplastic Agents; Area Under Curve; Biomarkers, Tumor; Carcinoembryonic Antigen; Drug Resistance, Neoplasm; Female; Humans; Kaplan-Meier Estimate; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Nucleosomes; Peptide Fragments; Phosphopyruvate Hydratase; Prognosis; Recombinant Proteins; ROC Curve; Sensitivity and Specificity; Small Cell Lung Carcinoma