Subtypes Of Lung Carcinoma Research Articles

Lung Cancer Management with Silibinin: A Historical and Translational Perspective.

The flavonolignan silibinin, the major bioactive component of the silymarin extract of Silybum marianum (milk thistle) seeds, is gaining traction as a novel anti-cancer therapeutic. Here, we review the historical developments that have laid the groundwork for the evaluation of silibinin as a chemopreventive and therapeutic agent in human lung cancer, including translational insights into its mechanism of action to control the aggressive behavior of lung carcinoma subtypes prone to metastasis. First, we summarize the evidence from chemically induced primary lung tumors supporting a role for silibinin in lung cancer prevention. Second, we reassess the preclinical and clinical evidence on the effectiveness of silibinin against drug resistance and brain metastasis traits of lung carcinomas. Third, we revisit the transcription factor STAT3 as a central tumor-cell intrinsic and microenvironmental target of silibinin in primary lung tumors and brain metastasis. Finally, by unraveling the selective vulnerability of silibinin-treated tumor cells to drugs using CRISPR-based chemosensitivity screenings (e.g., the hexosamine biosynthesis pathway inhibitor azaserine), we illustrate how the therapeutic use of silibinin against targetable weaknesses might be capitalized in specific lung cancer subtypes (e.g., KRAS/STK11 co-mutant tumors). Forthcoming studies should take up the challenge of developing silibinin and/or next-generation silibinin derivatives as novel lung cancer-preventive and therapeutic biomolecules.

A Bispecific DLL3/CD3 IgG-Like T-Cell Engaging Antibody Induces Antitumor Responses in Small Cell Lung Cancer.

Small cell lung cancer (SCLC) is the most lethal and aggressive subtype of lung carcinoma characterized by highly chemotherapy-resistant recurrence in the majority of patients. To effectively treat SCLC, we have developed a unique and novel IgG-like T-cell engaging bispecific antibody (ITE) that potently redirects T-cells to specifically lyse SCLC cells expressing Delta-like ligand 3 (DLL3), an antigen that is frequently expressed on the cell surface of SCLC cells, with no to very little detectable expression in normal tissues. The antitumor activity and mode of action of DLL3/CD3 ITE was evaluated in vitro using SCLC cell lines and primary human effector cells and in vivo in an SCLC xenograft model reconstituted with human CD3+ T-cells. Selective binding of DLL3/CD3 ITE to DLL3-positive tumor cells and T-cells induces formation of an immunological synapse resulting in tumor cell lysis and activation of T-cells. In a human T-cell engrafted xenograft model, the DLL3/CD3 ITE leads to an increase in infiltration of T-cells into the tumor tissue resulting in apoptosis of the tumor cells and tumor regression. Consistent with the mode of action, the DLL3/CD3 ITE treatment led to upregulation of PD-1, PD-L1, and LAG-3. This study highlights the ability of the DLL3/CD3 ITE to induce strictly DLL3-dependent T-cell redirected lysis of tumor cells and recruitment of T-cells into noninflamed tumor tissues leading to tumor regression in a preclinical in vivo model. These data support clinical testing of the DLL3/CD3 ITE in patients with SCLC.

Comparative proteogenomics profiling of non-small and small lung carcinoma cell lines using mass spectrometry.

BackgroundEvidences indicated that non-small-cell lung cancer (NSCLC) and small-cell lung cancer (SCLC) might originate from the same cell type, which however ended up to be two different subtypes of lung carcinoma, requiring different therapeutic regimens. We aimed to identify the differences between these two subtypes of lung cancer by using integrated proteome and genome approaches.Methods and MaterialsTwo representative cell lines for each lung cancer subtype were comparatively analysed by quantitative proteomics, and their corresponding transcriptomics data were obtained from the Gene Expression Omnibus database. The integrated analyses of proteogenomic data were performed to determine key differentially expressed proteins that were positively correlated between proteomic and transcriptomic data.ResultThe proteomics analysis revealed 147 differentially expressed proteins between SCLC and NSCLC from a total of 3,970 identified proteins. Combined with available transcriptomics data, we further confirmed 14 differentially expressed proteins including six known and eight new lung cancer related proteins that were positively correlated with their transcriptomics data. These proteins are mainly involved in cell migration, proliferation, and invasion.ConclusionThe proteogenomic data on both NSCLC and SCLC cell lines presented in this manuscript is complementary to existing genomic and proteomic data related to lung cancers and will be crucial for a systems biology-level understanding of the molecular mechanism of lung cancers. The raw mass spectrometry data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD015270.

Cytologic subtyping of lung carcinomas on Endobronchial ultrasound guided transbronchial needle aspiration (EBUS-TBNA) samples: Role of limited panel immunocytochemistry (ICC) as an adjunct to cytomorphology

Introduction: Majority of lung cancer presents at an advanced unresectable stage, emphasizing the diagnosis on cytological specimens like EBUS-TBNA. This study was aimed to subtype lung carcinomas on EBUS-TBNA specimens on smears and cell blocks (CB) and to assess the utility of limited panel ICC towards this subtyping. Materials and Methods: This is both retrospective and prospective study done on total of 142 cases for two and half years. The samples from either lung or mediastinal lymph nodes diagnosed as lung carcinoma on cytomorphology in correlation with clinico – radiological features were included. Extrapulmonary metastatic tumors, lymphomas and malignancies other than carcinomas were excluded. The cytologic material (EBUS-TBNA smears and CB) was obtained by EBUS TBNA procedure done on lung lesions and/or mediastinal lymph nodes and stained with required stains. All cases were first classified as small cell carcinoma (SCC) and non small cell carcinoma (NSCC) on cytomorphology as per 2004 WHO classification. NSCC was then further subtyped. ICC with TTF1/p63 antibodies was applied on NSCC and synaptophysin on SCC/neuroendocrine tumors. Napsin A was applied on p63/TTF1 negative cases. The results were expressed in frequencies and percentages. Result: On cytomorphology alone, 25 of 142 cases (17.6%) were classified as SCC and 117 cases (82.4%) as NSCC. NSCC was further subtyped on cytomorphology (smears+CB) into definitive (squamous cell carcinoma [SqCC]/adenocarcinoma [ADC]) v/s favouring (favouring SqCC/ ADC/ adenosquamous carcinoma) v/s not otherwise specified (NOS) in 41 (35.1%) v/s 33(28.2%) v/s 43(36.7%) cases respectively. Immunostains TTF1/ p63 and napsin A applied on possible 86 NSCC cases reduced NOS to 9.3 %. Synaptophysin confirmed all 20 cases of SCC and 1 large cell neuroendocrine carcinoma (LCNC). Overall in 142 cases, limited panel ICC reduced NSCC-NOS to 11.9%. Conclusion: Subtyping of lung carcinoma on EBUS- TBNA samples is feasible. Preparat

AN AUDIT OF LUNG CARCINOMAS AT A TERTIARY REFERRAL CENTRE

A generic diagnosis of ‘small cell or non-small cell carcinoma’ was sufficient in the past when there was no clinical demand for sub classification as it had no impact on therapy. Patients with advanced stage non-small cell lung carcinoma who received chemotherapy were typically treated with a ‘platinum doublet’ of cisplatin plus gemcitabine irrespective of histological subtype. At present, in contrast, sub classification of non-small cell lung carcinoma has significant treatment implications, especially for advanced stage tumors for which chemotherapy or targeted therapy is being considered. Consequently, pathologists are asked to subtype the tumor. The demand for sub typing is driven by oncologists, who now have several new targeted therapies, the efficacy of which varies by histological subtype.
 In view of demand of sub typing of lung carcinomas as well as the guidelines given by WHO 2015 classification which emphasises the importance of a separate classification for biopsies and cytology on one side and the resection specimen on the other, we have taken up this study.
 Keywords: Lung carcinoma, classification, core biopsies, immunohistochemistry

Current progress and future perspectives of polypharmacology : From the view of non-small cell lung cancer.

A pre-eminent subtype of lung carcinoma, Non-small cell lung cancer accounts for paramount causes of cancer-associated mortality worldwide. Undeterred by the endeavour in the treatment strategies, the overall cure and survival rates for NSCLC remain substandard, particularly in metastatic diseases. Moreover, the emergence of resistance to classic anticancer drugs further deteriorates the situation. These demanding circumstances culminate the need of extended and revamped research for the establishment of upcoming generation cancer therapeutics. Drug repositioning introduces an affordable and efficient strategy to discover novel drug action, especially when integrated with recent systems biology driven stratagem. This review illustrates the trendsetting approaches in repurposing along with their numerous success stories with an emphasize on the NSCLC therapeutics. Indeed, these novel hits, in combination with conventional anticancer agents, will ideally make their way the clinics and strengthen the therapeutic arsenal to combat drug resistance in the near future.

Identification of expression signatures for non-small-cell lung carcinoma subtype classification.

Non-small-cell lung carcinoma (NSCLC) mainly consists of two subtypes: lung squamous cell carcinoma (LUSC) and lung adenocarcinoma (LUAD). It has been reported that the genetic and epigenetic profiles vary strikingly between LUAD and LUSC in the process of tumorigenesis and development. Efficient and precise treatment can be made if subtypes can be identified correctly. Identification of discriminative expression signatures has been explored recently to aid the classification of NSCLC subtypes. In this study, we designed a classification model integrating both mRNA and long non-coding RNA (lncRNA) expression data to effectively classify the subtypes of NSCLC. A gene selection algorithm, named WGRFE, was proposed to identify the most discriminative gene signatures within the recursive feature elimination (RFE) framework. GeneRank scores considering both expression level and correlation, together with the importance generated by classifiers were all taken into account to improve the selection performance. Moreover, a module-based initial filtering of the genes was performed to reduce the computation cost of RFE. We validated the proposed algorithm on The Cancer Genome Atlas (TCGA) dataset. The results demonstrate that the developed approach identified a small number of expression signatures for accurate subtype classification and particularly, we here for the first time show the potential role of LncRNA in building computational NSCLC subtype classification models. The R implementation for the proposed approach is available at https://github.com/RanSuLab/NSCLC-subtype-classification.

Abstract 549: A novel T cell engaging bispecific antibody induces specific and efficacious lysis of small cell lung cancer cells in vitro and potent T cell re-directed anti-tumor activity in vivo

Abstract Small cell lung cancer (SCLC) is the most lethal and aggressive subtype of lung carcinoma characterized by highly chemotherapy resistant recurrence in the majority of patients. To effectively treat SCLC we have developed a unique T cell engaging bi-specific antibody that can potently re-direct T cells to specifically lyse SCLC cells expressing Delta-like Ligand 3 (DLL3). DLL3 is frequently expressed on the cell surface of neuroendocrine tumors, with no to very little expression in normal tissues, thus making DLL3 an ideal targeting antigen with which to safely re-direct cytolytic T cells to tumor cells. The bi-specific monovalent DLL3/CD3 IgG-like T cell Engager (ITE) is designed to have sustained serum exposure and simultaneously bind to CD3 on T cells and DLL3 on SCLC cells, resulting in formation of the cytolytic synapse. In vitro, unstimulated peripheral blood mononuclear cells (PBMCs) were co-cultured with several DLL3 expressing SCLC cell lines and increasing concentrations of the DLL3/CD3 ITE. The DLL3/CD3 ITE induced potent dose-dependent lysis of SCLC cell lines with EC90 values ranging from 15 to 150 ng/mL, whereas viability of DLL3-negative cells was unaffected, demonstrating the specificity of the DLL3/CD3 ITE for the DLL3 antigen. In addition, the DLL3/CD3 ITE induced DLL3-dependent activation and proliferation of T cells, and granzyme B and cytokine secretion. In vivo anti-tumor activity of the DLL3/CD3 ITE was assessed in NOG mice bearing subcutaneous xenografts of the SHP77 SCLC cell line and reconstituted with human T cells. Complete tumor regressions were observed with a dose of 0.25 mg/kg administered once weekly by i.v. administration, with the onset of activity being observed after the first dose. Consistent with the mode-of-action the DLL3/CD3 ITE led to a profound infiltration of both CD8 and CD4 T cell subsets in the tumors, which correlated with cleaved caspase 3 expression on tumor cells and tumor shrinkage. The inflammatory tumor microenvironment that was created by treatment with the DLL3/CD3 ITE also led to an increase of PD-1 on T cells and PD-L1 on the tumor cells. The DLL3/CD3 ITE shows cross-reactivity to both DLL3 and CD3 of human and cynomolgus monkey origin respectively, thus allowing the assessment of pharmacodynamics, pharmacokinetics, and safety in cynomolgus monkeys. In a single dose study in cynomolgus monkeys, the DLL3/CD3 ITE demonstrated antibody-like pharmacokinetic properties. These pre-clinical data demonstrate that the DLL3/CD3 ITE is a highly potent, efficacious, and DLL3-selective T cell redirecting agent, and supports future clinical development. Citation Format: Susanne Hipp, Vladimir Voynov, Barbara Drobits-Handl, Francesca Trapani, Craig Giragossian, Justin M. Scheer, Paul J. Adam. A novel T cell engaging bispecific antibody induces specific and efficacious lysis of small cell lung cancer cells in vitro and potent T cell re-directed anti-tumor activity in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 549.

Histopathologic Analysis of Lung Cancer Incidence Associated with Radon Exposure among Ontario Uranium Miners.

Although radon is a well-established contributor to lung cancer mortality among uranium miners, the effects of radon decay products on different histopathologies of lung carcinoma are not well established. Using a retrospective cohort design, this study aims to examine the risks of lung cancer by histological subtypes associated with exposure to radon decay products among the Ontario Uranium Miners cohort. Cases were stratified by histological groups, and associated risks were estimated for cumulative radon exposure after adjustment for attained age and calendar period. Between 1969 and 2005, 1274 incident cases of primary lung cancer were identified. Of these, 1256 diagnoses (99%) contained information on histology. Squamous cell carcinoma was most common (31%), followed by adenocarcinoma (20%), large cells (18%), small cell lung carcinoma (14%), and other or unspecified cell types (17%). Of the histological sub-groups, small cell lung carcinoma had the strongest association with cumulative radon exposure; compared to the reference group (<1 cumulative working level months (WLM)), the highest exposure category (>60 cumulative WLM) had a relative risk (RR) of 2.76 (95% CI: 1.67–4.57). Adenocarcinoma had the lowest risk and was not significantly associated with exposure to radon decay products (RR = 1.49, 95% CI: 0.96–2.31). An increasing, linear trend in relative risk was noted with increasing cumulative WLM across small cell, squamous cell, and large cell lung carcinomas (Ptrend < 0.05). Similarly, the excess relative risk (ERR) per WLM was highest for small cell lung carcinoma (ERR/WLM = 0.15, p < 0.01), followed by squamous cell carcinoma (ERR/WLM = 0.12, p < 0.01). Non-statistically significant excess risk was observed for adenocarcinoma (ERR/WLM = 0.004, p = 0.07). Our analysis of the Ontario Uranium Miners cohort data shows differences in the magnitude of the risks across four histological subtypes of lung carcinoma; the strongest association was noted for small cell lung carcinoma, followed by squamous cell, large cell, and lastly adenocarcinoma, which showed no significant associations with exposure to radon decay products.

Intra-tumoural heterogeneity characterization through texture and colour analysis for differentiation of non-small cell lung carcinoma subtypes

Radiomics has shown potential in disease diagnosis, but its feasibility for non-small cell lung carcinoma (NSCLC) subtype classification is unclear. This study aims to explore the diagnosis value of texture and colour features from positron emission tomography computed tomography (PET-CT) images in differentiation of NSCLC subtypes: adenocarcinoma (ADC) and squamous cell carcinoma (SqCC). Two patient cohorts were retrospectively collected into a dataset of 341 18F-labeled 2-deoxy-2fluoro-d-glucose ([18F] FDG) PET-CT images of NSCLC tumours (125 ADC, 174 SqCC, and 42 cases with unknown subtype). Quantification of texture and colour features was performed using freehand regions of interest. The relation between extracted features and commonly used parameters such as age, gender, tumour size, and standard uptake value (SUVmax) was explored. To classify NSCLC subtypes, support vector machine algorithm was applied on these features and the classification performance was evaluated by receiver operating characteristic curve analysis. There was a significant difference between ADC and SqCC subtypes in texture and colour features (P < 0.05); this showed that imaging features were significantly correlated to both SUVmax and tumour diameter (P < 0.05). When evaluating classification performance, features combining texture and colour showed an AUC of 0.89 (95% CI, 0.78–1.00), colour features showed an AUC of 0.85 (95% CI, 0.71–0.99), and texture features showed an AUC of 0.68 (95% CI, 0.48–0.88). DeLong’s test showed that AUC was higher for features combining texture and colour than that for texture features only (P = 0.010), but not significantly different from that for colour features only (P = 0.328). HSV colour features showed a similar performance to RGB colour features (P = 0.473). The colour features are promising in the refinement of NSCLC subtype differentiation, and features combining texture and colour of PET-CT images could result in better classification performance.

O impacto da histologia do carcinoma pulmonar na frequência das metástases ósseas

ObjectivesLung cancer is the leading cause of oncological deaths, and bones are one of the most common sites of metastasis from this cancer type. The objective of this study was to evaluate the influence of lung carcinoma histology on the frequency of bone metastases. MethodsThis retrospective study evaluated medical records from 407 patients who were diagnosed with lung carcinoma between 2003 and 2012. The prevalence of bone metastases and their association with histological subtypes were evaluated using the chi‐squared test, odds ratios (ORs), and 95% confidence intervals (CIs). Overall survival was evaluated using the Kaplan‐Meier method. ResultsThe prevalence of bone metastases was 28.2% (n=115) and the spine was the most frequent site (98 metastases; 32.1%). Adenocarcinoma was the most common histological subtype of lung carcinoma (46.7%), and was significantly more frequent among patients with bone metastases (58.3% vs. 42.1%; p=0.003; OR=1.92; 95% CI:1.29–2.97). Squamous cell carcinoma was significantly less frequent among patients with bone metastases (13.0% vs. 29.8%; p=0.0004; OR=0.35; 95% CI: 0.19–0.64). The median survival time after the first bone metastasis diagnosis was four months. ConclusionAdenocarcinoma was the most common histological subtype of lung carcinoma and was significantly associated with a higher risk of developing bone metastases.

Human epididymis protein 4 may not be a reliable screening biomarker for detecting lung carcinoma patients.

Human epididymis protein 4 (HE4) acts as a protease inhibitor. It has been detected in the serum of patients with lung cancer patients and its utility for lung cancer screening was found in different studies. Nevertheless, little is known regarding the expression of HE4 in lung carcinoma subtypes. The aim of the present study was to investigate whether HE4 expression is a reliable marker for detecting any lung carcinoma subtypes, including small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC) and adenocarcinoma (AC). In total, 141 lung carcinoma patients were enrolled in the study. Biopsy samples were obtained from bronchoscopic biopsy. The tumors were classified as SCLC (group 1, 54 cases) or NSCLC (group 2, 87 cases) based on histology and immunohistochemistry. The latter was sub-grouped as adenocarcinoma (group 2a, AC) and squamous cell carcinoma (group 2b, SCC). The immunohistochemical expression of HE4 was compared between the groups. The study revealed that the majority of the SCLC and SCC cases were devoid of HE4 (90.1 and 89.65%, respectively). Approximately 10% of cases had HE4 immune expression and the staining was focal and moderate throughout the tumor tissue. On the other hand, 78.8% of AC expressed HE4 and the staining was diffuse and strong. The overall HE4 expression in the lung cancer patients was 33.7%. In conclusion, the results of the present study have shown that HE4 is expressed in adenocarcinoma of the lung but it is not frequent in SCC and SCLC. The value of HE4 as a screening biomarker for lung cancer is limited but its presence exclusively in adenocarcinoma may provide new insight for targeted therapy.

The diagnostic value of TTF1, P63, HMWK [34βE12], CK7, and CD56 immunostaining in classification of lung carcinoma

The histologic distinction of small cell from non-small cell lung carcinoma and correct identification of all subtypes of lung carcinoma are very important in treatment management. The main method for histologic classification of lung tumors is based on morphology. However, in small bronchoscopic biopsies in particular, distinction is very difficult upon morphology alone. The current study aimed at evaluating the utility of a panel of antibodies, consisting of thyroid transcription factor (TTF-1), P63, high molecular weight keratin [HMWK (34βE12)], cytokeratin (CK7), and cluster of differentiation (CD56) for accurate distinction of bronchogenic carcinomas. Bronchoscopic biopsies of 60 lung carcinoma cases including 20 small cell carcinomas, 20 adenocarcinomas, and 20 squamous cell carcinomas (SCCs) with typical morphologic features were selected. All these cases were immunohistochemically stained for TTF-1, P63, HMWK (34βE12), CK7, and CD56. All immunostained slides were scored as either positive or negative. The mean age of the patients was 60 years; ranged from 35 to 81. Sixteen patients were female and 44 were male. All adenocarcinomas were positive for CK7 and most of them (18/20; 90%) were positive for TTF-1. Most of small cell lung carcinomas were positive for TTF-1 (17/20; 85%), and CD56 (18/20; 90%). All squamous cell carcinomas (SCCs) were negative for TTF-1, but most of them were positive for HMWK (34βE12) and P63. The obtained data showed that TTF-1, P63, CK7, CD56 and/or 34βE12 represent a useful panel of antibodies to identify lung carcinoma subtypes in small bronchoscopicbiopsies.

Targeted therapy in small cell lung cancer: can DLL3 notch up a victory?

Small cell lung cancer (SCLC) is the most lethal and aggressive subtype of lung carcinoma, responsible for ~13–18% of lung cancer death with no appreciable improvements in outcomes or treatment options for the last 30 years. The clinical behavior of SCLC is tailor made for nihilism with excellent initial overall response rates transforming to inevitable chemotherapy resistant recurrence in the majority of patients. Targeted therapies to date have failed with little to no efficacy in unselected populations. Naturally, this state of affairs has led to an underfunded SCLC research community, and historical pharmaceutical disinterest in this “graveyard of drug development”. The National Cancer Institute (NCI) and worldwide refocus upon “recalcitrant” carcinomas has led to renewed interest in SCLC making this the perfect opportunity to consider how and why targeted therapy in unselected SCLC has failed so consistently. The critical factors are both biological factors and structural limitations to previous targeted therapy studies in SCLC.

Utility of Cluster of Differentiation 5 and Cluster of Differentiation 117 Immunoprofile in Distinguishing Thymic Carcinoma from Pulmonary Squamous Cell Carcinoma: A Study on 1800 Nonsmall Cell Lung Cancer Cases.

Background:Mediastinum harbors a mixed bag of neoplastic lesions with varied therapeutic and prognostic implications. Distinguishing pulmonary squamous cell carcinoma from thymic carcinoma is a challenging task owing to rather nonspecific clinicoradiological features and a considerable overlap in histomorphological features. Thus, we evaluated the diagnostic utility of cluster of differentiation 5 (CD5) and CD117 immunohistochemical markers in distinguishing these tumors.Materials and Methods:The archives of department of histopathology were retrospectively reviewed from June 2012 to May 2016. Formalin-fixed paraffin-embedded blocks of 1800 cases diagnosed as nonsmall cell lung carcinoma were retrieved, and immunohistochemical expression of CD5 and CD117 was evaluated in these cases.Results:Adenocarcinoma (980; 54.44%) was the most common histological subtype of lung carcinoma observed in our study. CD117 was positive in 171 out of 1800 cases (9.5%) of which 120 cases (70.17%) were adenocarcinoma, followed by 40 cases (23.4%) of squamous cell carcinoma. Immunoreactivity for CD5 was observed in 209 cases (11.61%), 200 (95.7%) cases of which were diagnosed as adenocarcinoma. None of the cases diagnosed as squamous cell carcinoma on histomorphology showed CD5 immunoexpression.Conclusion:While thymic squamous cell carcinomas are well known for CD5 and CD117 coexpression, none of the cases of squamous cell carcinoma arising in lung express CD5. These markers are a diagnostic tool to distinguish a primary lung squamous cell carcinoma from thymic carcinoma, particularly in the setting of a central pulmonary lesion with mediastinal involvement.

Histologic Subtyping of Lung Carcinomas: From The Targeted Therapy Perspective

Histologic Subtyping of Lung Carcinomas: From The Targeted Therapy Perspective

Diagnostic concordance of non-small cell lung carcinoma subtypes between biopsy and cytology specimens obtained during the same procedure.

The objectives of this study were: 1) to determine the diagnostic concordance of non-small cell lung carcinoma (NSCLC) subtypes in cytology and biopsy specimens taken during the same procedure and evaluate the causes of discordance; and 2) to determine the frequency of immunohistochemistry (IHC) use for subtyping NSCLC. Biopsy and cytology specimens that were obtained at the same procedure and diagnosed as NSCLC between January 2011 and December 2014 at the McGill University Health Center were identified (n = 226 pairs). The diagnostic concordance between the 2 methods was evaluated. The slides from discordant cases were reviewed, and final diagnoses were made based on IHC, resection specimens, or pathologist discussion. Concordance in subtype diagnosis was perfect (adeno-adeno or squamous-squamous) in 66.2% of cases and was partial (adeno or squamous vs non-small cell) in 23%; discordance (adeno vs squamous) was observed in 7.8%. Although subtyping was not possible (ie, the final diagnosis was NSCLC, not otherwise specified) in 12.8% of biopsy specimens and 16.3% of cytology specimens, specific subtyping was not achieved in only 3% of cases when both modalities were considered. IHC was used in 47% of biopsy cases and 13% of cytology cases. Subtyping of NSCLC can be achieved in most cases (97%) by considering findings in both biopsy and cytology specimens, and concordance in subtyping between cytology and biopsy specimens can be reached in a high percentage of cases (89.2%). Cancer Cytopathol 2016;124:737-43. © 2016 American Cancer Society.

Selective expression of transthyretin in subtypes of lung cancer.

Transthyretin (TTR) is expressed primarily in liver, choroid plexus of brain and pancreatic islet A and B cells. It is also synthesized in some endocrine tumors. In the present study, the protein expression of TTR in lung cancer tissues and cell lines was investigated by western blot. The mRNA expression of TTR in 24 pairs of frozen lung cancer tissues was examined by RT-PCR. The specific expression and cellular distribution of TTR were also evaluated in 104 paraffin-embedded lung cancer samples and 3 normal lung tissues by immunohistochemistry. Similarly, the subcellular localization and expression of TTR were further analyzed in lung cancer cell lines. With the exception of mucinous adenocarcinoma, the expression of TTR protein was observed in all tested subtypes of lung carcinoma. Adenocarcinoma displayed the highest positive expression rate of TTR, accounting for 84.4%, and the positive expression rate of TTR was up to 85.7% at stages III and IV. The secretory bubbles with strong TTR staining were observed in luminal cells of lung cancer. Furthermore, the localization of TTR in the cytoplasm of lung cancer cells and the secretion of TTR into extracellular milieu were also confirmed. Taken together, TTR is selectively synthesized in lung cancer cells and can be secreted extracellularly.

The Expression of miR-375 Is Associated with Carcinogenesis in Three Subtypes of Lung Cancer.

Many studies demonstrated unique microRNA profiles in lung cancer. Nonetheless, the role and related signal pathways of miR-375 in lung cancer are largely unknown. Our study investigated relationships between carcinogenesis and miR-375 in adenocarcinoma, squamous cell carcinoma and small cell lung carcinoma to identify new molecular targets for treatment. We evaluated 723 microRNAs in microdissected cancerous cells and adjacent normal cells from 126 snap-frozen lung specimens using microarrays. We validated the expression profiles of miR-375 and its 22 putative target mRNAs in an independent cohort of 78 snap-frozen lung cancer tissues using quantitative reverse-transcriptase PCR. Moreover, we performed dual luciferase reporter assay and Western blot on 6 targeted genes (FZD8, ITGA10, ITPKB, LRP5, PIAS1 andRUNX1) in small cell lung carcinoma cell line NCI-H82. We also detected the effect of miR-375 on cell proliferation in NCI-H82. We found that miR-375 expression was significantly up-regulated in adenocarcinoma and small cell lung carcinoma but down-regulated in squamous cell carcinoma. Among the 22 putative target genes, 11 showed significantly different expression levels in at least 2 of 3 pair-wise comparisons (adenocarcinoma vs. normal, squamous cell carcinoma vs. normal or small cell lung carcinoma vs. normal). Six targeted genes had strong negative correlation with the expression level of miR-375 in small cell lung carcinoma. Further investigation revealed that miR-375 directly targeted the 3’UTR of ITPKB mRNA and over-expression of miR-375 led to significantly decreased ITPKB protein level and promoted cell growth. Thus, our study demonstrates the differential expression profiles of miR-375 in 3 subtypes of lung carcinomas and finds thatmiR-375 directly targets ITPKB and promoted cell growth in SCLC cell line.

Trefoil factor 3 as a novel biomarker to distinguish between adenocarcinoma and squamous cell carcinoma.

In carcinoma, such as of the lung, the histological subtype is important to select an appropriate therapeutic strategy for patients. However, carcinomas with poor differentiation cannot always be distinguished on the basis of morphology alone nor on clinical findings. Hence, delineation of poorly differentiated adenocarcinoma and squamous cell carcinoma, the 2 most common epithelial-origin carcinomas, is pivotal for selection of optimum therapy. Herein, we explored the potential utility of trefoil factor 3 (TFF3) as a biomarker for primary lung adenocarcinoma and extrapulmonary adenocarcinomas derived from different organs. We observed that 90.9% of lung adenocarcinomas were TFF3-positive, whereas no expression of TFF3 was observed in squamous cell carcinomas. The subtype of lung carcinoma was confirmed by four established biomarkers, cytokeratin 7 and thyroid transcription factor 1 for adenocarcinoma and P63 and cytokeratin 5/6 for squamous cell carcinoma. Furthermore, expression of TFF3 mRNA was observed by quantitative PCR in all of 11 human lung adenocarcinoma cell lines and highly correlated with markers of the adenocarcinomatous lineage. In contrast, little or no expression of TFF3 was observed in 4 lung squamous cell carcinoma cell lines. By use of forced expression, or siRNA-mediated depletion of TFF3, we determined that TFF3 appeared to maintain rather than promote glandular differentiation of lung carcinoma cells. In addition, TFF3 expression was also determined in adenocarcinomas from colorectum, stomach, cervix, esophagus, and larynx. Among all these extrapulmonary carcinomas, 93.7% of adenocarcinomas exhibited TFF3 positivity, whereas only 2.9% of squamous cell carcinomas were TFF3-positive. Totally, 92.9% of both pulmonary and extrapulmonary adenocarcinomas exhibited TFF3 positivity, whereas only 1.5% of squamous cell carcinomas were TFF3-positive. In conclusion, TFF3 is preferentially expressed in adenocarcinoma and may function as an additional biomarker for distinguishing adenocarcinoma from squamous cell carcinoma.