Large Cell Carcinoma Research Articles

Features of the expression of prognostic markers p16, Her2 / new and changes in the corresponding genes in individual phenotypes of cancers of unknown primary origin.

Background. Approximately 2/3 of cancers of unknown primary origin (CUP) are adenocarcinomas with mucin production, the formation of gland-like structures and immunohistochemistry results that can determine the histological subtype, but cannot determine the exact primary place of origin. The remaining 1/3 of CUP is a mixture of non-adenocarcinomas, including squamous cell carcinomas, neuroendocrine carcinomas, small-cell and large-cell undifferentiated carcinomas. Objective. In such cases, molecular profile identification and gene sequencing may be a promising approach to the classification and direct treatment of CUP. Methods. A retrospective analysis of biopsy or postoperative material of 61 patients (34 women and 27 men) aged 26 to 74 years (mean 53.4 ± 11.42; median 52) with an initial diagnosis of CUP was performed. Immunohistochemical studies were carried out according to the protocols of TermoScientific (USA) with antibodies to p16INK4a and HER2/neu. Fluorescence in situ hybridization was performed according to the protocol of ZytoVision GmbH (Germany). Changes in the ERBB2 and CDKN2A genes were evaluated for gene amplification and deletion of the 9p21 locus, respectively, with double-labeled DNA probes. Results. Of the 61 studied cases of CUP, p16 expression was positive in 31 cases (51%), the basis for which was 9 homozygous and 19 heterozygous 9p21 deletions, which were found in all but one of the studied CUP phenotypes. HER2 / neu expression was positive (2 + / 3 +) in 15 of 61 (25%) CUP, but only in 9 of them had actual amplification of the ERBB2 gene on FISH in 5 phenotypes. Conclusion. The reduced number of phenotypes with amplification of the ERBB2 gene makes it more specific for the differential diagnosis of CUP, compared with the deletion of 9p21 (p16 / CDKN2A), which is likely to be universal in acquiring an aggressive course of carcinomas of various localizations.

Inhibition of bone morphogenetic protein signaling reduces viability, growth and migratory potential of non-small cell lung carcinoma cells.

BMP signaling has an oncogenic and tumor-suppressing activity in lung cancer that makes the prospective therapeutic utility of BMP signaling in lung cancer treatment complex. A more in-depth analysis of lung cancer subtypes is needed to identify BMP-related therapeutic targets. We sought to examine the influence of BMP signaling on the viability, growth and migration properties of the cell line LCLC-103H, which originates from a large cell lung carcinoma with giant cells and an extended aneuploidy. We used BMP-4 and LDN-214117 as agonist/antagonist system for the BMP receptor type I signaling. Using flow cytometry, wound healing assay, trans-well assay and spheroid culture, we examined the influence of BMP signaling on cell viability, growth and migration. Molecular mechanisms underlying observed changes in cell migration were investigated via gene expression analysis of epithelial-mesenchymal transition (EMT) markers. BMP signaling inhibition resulted in LCLC-103H cell apoptosis and necrosis 72h after LDN-214117 treatment. Cell growth and proliferation are markedly affected by BMP signaling inhibition. Chemotactic motility and migratory ability of LCLC-103H cells were clearly hampered by LDN-214117 treatment. Cell migration changes after BMP signaling inhibition were shown to be coupled with considerable down-regulation of transcription factors involved in EMT, especially Snail. BMP signaling inhibition in LCLC-103H cells leads to reduced growth and proliferation, hindered migration and accelerated cell death. The findings contribute to the pool of evidence on BMP signaling in lung cancer with a possibility of introducing BMP signaling inhibition as a novel therapeutic approach for the disease.

Pulmonary carcinosarcoma: analysis from the Surveillance, Epidemiology and End Results database.

Pulmonary carcinosarcoma (PCS) is a rare neoplasm. This study explored the clinicopathological characteristics and survival outcomes of PCS. The Surveillance, Epidemiology and End Results (SEER) database (1988-2014) was queried for PCS. Overall survival (OS) was evaluated by multivariable Cox regression and nomograms were constructed to predict 3-year OS for PCS. Prognostic performance was evaluated using concordance index and area under the curve analysis. In M0 surgically treated patients, interaction assessments were performed using likelihood ratio tests. Subgroup analysis was performed according to patient age. The clinical features of PCSs were further compared to other non-small-cell lung cancers (NSCLCs). Multivariable analysis identified age [hazard ratio (HR) 1.03, 95% confidence interval (CI) 1.01-1.04], surgery (HR 0.53, 95% CI 0.36-0.77) and chemotherapy (HR 0.51, 95% CI 0.36-0.73) as significantly associated with OS. The nomogram had a concordance index of 0.747 and an area under the curve of 0.803. The association between age and OS was stronger in those receiving pneumonectomy (P = 0.04 for interactions) compared to those that did not (HR 5.14, 95% CI 1.64-16.07), and was associated with a poorer outcome compared to lobectomy amongst the elderly (age ≥ 70 years). Patients with PCS were more likely to receive surgical treatment and had lower lymphatic metastasis compared to adenocarcinoma, squamous cell carcinoma and large cell carcinoma (all P < 0.05). PCS had unique clinical features compared to common types of NSCLCs in terms of lymphatic invasion and surgical treatment. Pneumonectomy was associated with poorer survival in elderly patients.

P14.97 High risk of venous thromboembolism in patients with brain metastases from non-small cell lung cancer

Abstract BACKGROUND Venous thromboembolism (VTE) is a common complication in patients with cancer. In general, patients with metastatic disease are at highest risk. Lung cancer belong to those tumor entities with a particularly high risk of VTE, ranging between 3–13.8%. However, little is known about the VTE rate in lung cancer patients with brain metastases. MATERIAL AND METHODS Our study was conducted in the framework of the Vienna Brain Metastasis Registry. Clinical data and VTE events during the course of the disease were recorded via retrospective chart review. In this analysis, non-small cell lung cancer (NSCLC) patients with a resection of brain metastases at the Medical University of Vienna between 2006 and 2010 were included. RESULTS In total, 69 NSCLC patients with brain metastases were analyzed. Overall, 69.6% (48/69) patients had an adenocarcinoma, 13% (9/69) a squamous cell carcinoma, 8.7% (6/69) a large cell carcinoma and 8.7% (6/69) other primary tumor histologies. After cancer diagnosis, 20.3% (14/69) patients developed VTE during the course of the disease. Of those, 85.7% (12/14) thromboembolic events occurred after the diagnosis of brain metastases. CONCLUSION Based on our data, patients with brain metastases from NSCLC have a very high VTE risk. Further investigations are needed in order to identify patients with distinct VTE risk profiles. Patients at high risk might potentially benefit from primary thromboprophylaxis over the high risk of intracerebral bleeding.

The Prevalence of PIK3CA Biomarker in Tumor Microenvironment of Human non-small Cell Lung Cancer

Background. Phosphatidyl 3-kinases (PI3K) are a family of lipid kinases involved in many cellular processes, including cell growth, proliferation, differentiation, motility, and survival, that can inhibit therapy profoundly. NSCLC accounts for approximately 80% of all lung cancers and is further subtyped into adenocarcinoma, squamous cell carcinoma, and large cell carcinoma. Somatic oncogenic alterations can further molecularly subdivide these NSCLC subtypes. The general objective of the current work was to study the prevalence of PIK3CA expression and mutations in tumor microenvironment of human NSCLC. Materials and Methods. Thirty-three paraffin blocks were utilized from NSCLC archival cases for further investigations by using immunohistochemical staining and PCR/sequencing techniques for examination of PI3KCA cellular expression and detection of mutation in exons 9 and / or 20 of PI3KCA, respectively. Results. Immunohistochemical results showed that PIK3CA protein was localized in the cytoplasm of NSCLC as brown granules. The rating of scores of PIK3CA immunostaining of archived NSCLC revealed that twenty six cases (78.8%) showed positive immunohistochemical staining while seven cases (21.2%) showed negative immune staining. PCR/sequencing results of exon 9 revealed six substitution mutations and three indels (insertion / deletion) mutations were observed in the alignment. It was found that all the observed SNP positions were novel except 74681. The novelty of each observed SNP was detected using the NCBI SNP detector and it was found that all the observed SNP positions were novel except 7468. PCR/sequencing results of exon 20 revealed that the alignment results of all the patient samples for exon 20 revealed the presence of SNPs in variants no. 3, 5, and 7. All of these variations were only substitution mutations of this PCR amplicons. The exact position of each observed mutation was mentioned in the NCBI reference sequences. Conclusion. The data in hands shows that PIK3CA-mutated NSCLC is a clinically and genetically heterogeneous group. This holds true for the AD and SQCC subgroups and strongly suggests that PIKCA mutations do not define a distinct lung cancer subgroup amendable to specific therapy. Rather, these mutations seem to represent passenger mutations widely distributed among the other genetically defined subgroups. The status PIK3CA mutation may serve as a prognostic factor of poor survival.

Stereotactic Body Radiotherapy for Large Primary Renal Cell Carcinoma: A Report from the International Radiosurgery Oncology Consortium for Kidney (IROCK)

Thermal ablation is an established option for small renal cell carcinomas (RCC). However, patients with larger (T1b, >4cm) RCC not suitable for surgery have few treatment options as thermal ablation is less effective in this setting. We hypothesize that SBRT has similar local efficacy (>90%) in larger RCC to that of small RCC. Individual patient data from nine IROCK institutions in Germany, Australia, USA, Canada, and Japan were pooled. Outcomes from larger RCC were compared to small RCC. Patients with metastatic disease or urothelial histology were excluded from the database (n=31). Demographics, treatment, oncologic and renal function outcomes were compared between cohorts using the Chi-square test, Fisher’s exact test, two-sample T-test, or Wilcoxon rank sum test as appropriate. Kaplan-Meier estimates and univariable and multivariable Cox proportional hazards regression were generated for oncologic outcomes. There were 95 patients with larger RCC identified from 192 eligible patients. Median follow up was 2.5 years. Median tumor size in the larger RCC group was 4.9 cm compared to 2.9 cm in the small RCC group. In the overall cohort, normal renal function (eGFR>90mL/min) at baseline was recorded in 9.0% of patients, with 26% having moderate to severe dysfunction (eGFR <45mL/min). Patients with large RCC were of poorer performance status (ECOG 0-1 in 81.1% vs. 93.8%, p = 0.008) and were older (mean age: 75 years vs. 69 years, p < 0.001). Toxicities ≥ grade 2 occurred in 12 patients (6.3%) with no differences in toxicity rates between cohorts (p = 0.526). Median baseline eGFR was lower in the larger RCC cohort (59.0mL/min vs. 68.5 mL/min, p = 0.013), and demonstrated a median reduction post-SBRT of -5.8 v mL/min versus -2.9 mL/min, p = 0.073). There were 8 recurrences (8.4%) in larger RCCs, compared to 1 recurrence (1.0%) in the small RCC cohort. Local control at 1-year was 100% in both cohorts, and at 3-years was similar at 96.6% vs. 98.5%, respectively (log-rank p = 0.512). Distant control was worse in larger RCC at 1- and 3-years, at 97.8% vs. 100% and 87.0% vs. 100% respectively (log-rank p = 0.006). Cancer-specific survival (CSS) at 3-years was 91.4% versus 96.2% (log-rank p = 0.224). Overall survival (OS) at 3-years was 72.3% vs. 84.6% (log-rank p = 0.294). On multivariable analysis, higher pre-SABR creatinine (hazard ratio [HR] per 10 μmol/L: 1.08, p = 0.002) and poor performance status (HR: 3.85, p = 0.004) were associated with inferior OS. Increasing tumor size (HR per 1 cm: 1.18, p = 0.028) was associated with inferior CSS. SBRT for larger RCC appears tolerable and similarly locally effective as in small RCC, although associated with a higher likelihood of metastatic progression and death. Efforts to reduce risk of distant recurrence, such as adjuvant systemic therapy, should be further investigated.

Primary Laryngeal Neuroendocrine Carcinoma: Two Cases of an Unusual Tumor in a Rare Location

Introduction: Neuroendocrine neoplasm is an infrequent tumor in the larynx which can be sub-classified into five different types: typical carcinoid, atypical carcinoid, large cell carcinoma, small cell carcinoma, and paraganglioma. This tumor mostly occurs in smoker men. The common location of neuroendocrine tumor of the larynx is in the supraglottic area. Because of the complex nature of neuroendocrine neoplasms of the larynx, we found it so interesting to present. We emphasized the clinical importance and aggressive nature of this tumor despite its rarity. Case Presentation: In the present study, we presented 2 cases of atypical carcinoid in non-smoker females. Case 1 was a 74-year-old non-smoker woman who was referred with a history of chronic dysphagia and left otalgia since one year ago. Laryngoscopy revealed a nonulcerated supraglottic epitheliomatous lesion. Biopsy showed atypical carcinoid. The patient had no lymphadenopathy or metastasis in the further workup. The second case was a 60-year-old non-smoker woman with a 4-month history of hoarseness, odynophagia accompanied by cervical lymphadenopathy. Laryngoscopy showed a smooth surface non-ulcerated transglottic tumor. Right vocal cord mobility was restricted. Conclusions: Atypical carcinoid tumor is an underdiagnosed laryngeal malignancy. Although neuroendocrine tumor of the larynx is rare, physicians should be familiar with its unspecified presentations, pathology, clinical course, and prognosis.

CYFRA21-1 tests in the diagnosis of non-small cell lung cancer: A meta-analysis.

The aim of the study was to evaluate the diagnostic value of soluble fragment of cytokeratin 19 (CYFRA21-1) tests in detecting non-small cell lung cancer (NSCLC), including squamous cell carcinoma, lung adenocarcinoma, and large cell carcinoma. The relevant studies were identified from PubMed, Embase and the Cochrane Library before November 2018. Summary estimates for sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio of CYFRA21-1 tests for the diagnosis of NSCLC were calculated using the random effects model. A summary receiver operating characteristic (SROC) curve was used to assess the overall effectiveness of the test. Meta-DiSc 1.4 and Stata11.0 were applied to the statistical analysis. Publication bias was detected using Egger's test. A total of 22 studies consisting of 7910 NSCLC patients (squamous cell carcinoma/lung adenocarcinoma/large cell carcinoma) and 2630 benign lesions patients that met the inclusion criteria were included. The meta-analysis showed that CYFRA21-1 tests had a relatively high accuracy for squamous cell carcinoma detection and a lower accuracy for lung adenocarcinoma detection. The overall sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio of CYFRA21-1 tests for squamous cell carcinoma detection were 0.72 (95% confidence interval (CI) 0.70, 0.74), 0.94 (95% CI 0.92, 0.95), 9.73 (95% CI 7.06, 13.40), 0.37 (95% CI 0.29, 0.47), and 27.30 (95% CI 17.68, 42.16), respectively. The area under the SROC curve was 0.9171 (Q* = 0.8500). No publication bias was tested in the squamous cell carcinoma (P = 0.567) and lung adenocarcinoma (P = 0.378) groups. CYFRA21-1 tests might be appropriate for detecting squamous cell carcinoma.

Surgical complexity and prognostic outcome of small volume renal cell carcinoma with high-level venous tumor thrombus and large volume renal cell carcinoma with low-level thrombus.

Background:Radical nephrectomy with thrombectomy is one of the most difficult and complicated urological operations. But the roles of renal tumor volume and thrombus level in surgical complexity and prognostic outcome are not clear. This study aimed to evaluate the surgical complexity and prognostic outcome between the volume of renal cell carcinoma (RCC) and the level of venous tumor thrombus.Methods:The clinical data of 67 RCC cases with renal vein or inferior vena cava (IVC) tumor thrombus from January 2015 to May 2018 were retrospectively analyzed. Among these 67 cases, 21 (31.3%) were small tumors with high-level thrombus (tumor ≤7 cm in diameter and thrombus Neves Level II–IV), while 46 (68.7%) were large tumors with low-level thrombus group (tumor >7 cm in diameter and thrombus Level 0–I). Clinical features, operation details, and pathology data were collected. Univariable and multivariable logistic regression analyses were applied to evaluate the risk factors for small tumor with high-level thrombus.Results:Patients with small tumors and high-level thrombus were more likely to have longer operative time (421.9 ± 135.1 min vs. 282.2 ± 101.9 min, t = 4.685, P < 0.001), more surgical bleeding volume (1200 [325, 2900] mL vs. 500 [180, 1000] mL, U = 270.000, P = 0.004), more surgical blood transfusion volume (800 [0, 1400] mL vs. 0 [0, 800] mL, U = 287.500, P = 0.004), more plasma transfusion volume (0 [0, 800] mL vs. 0 [0, 0] mL, U = 319.000, P = 0.004), higher percentage of open operative approach (76.2% vs. 32.6%, χ2 = 11.015, P = 0.001), higher percentage of IVC resection (33.3% vs. 0%, χ2 = 17.122, P < 0.001), and higher percentage of post-operative complications (52.4% vs. 19.6%, χ2 = 7.415, P = 0.010) than patients with large tumors and low-level thrombus. In multivariate analysis, decreased hemoglobin (Hb) (odds ratio [OR]: 0.956, 95% confidence interval [CI]: 0.926–0.986, P = 0.005) and non-sarcomatoid differentiation (OR: 0.050, 95% CI: 0.004–0.664, P = 0.023) were more likely to form small tumors with high-level tumor thrombus rather than large tumor with small tumor thrombus. The estimated mean cancer-specific survival times of small tumor with high-level thrombus and large tumor with low-level thrombus were 31.6 ± 3.8 months and 32.5 ± 2.9 months, without statistical significance (P = 0.955). After univariate and multivariate Cox proportional hazard survival regression analyses, only distant metastasis (hazard ratio [HR]: 3.839, P = 0.002), sarcomatoid differentiation (HR: 7.923, P < 0.001), alkaline phosphatase (HR: 2.661, P = 0.025), and severe post-operative complications (HR: 10.326, P = 0.001) were independent predictors of prognosis.Conclusions:The level of the tumor thrombus was more important than the diameter of the primary kidney tumor in affecting the complexity of surgery. In the same T3 stage, neither the renal tumor diameter nor the tumor thrombus level was an independent risk factor for prognosis.

Prognostic significance of the pattern of pathological N1 lymph node metastases for non-small cell lung cancer.

In patients with non-small cell lung cancer (NSCLC) the pathologic lymph node status N1 is a heterogeneous entity, and different forms of lymph node involvement may represent different prognoses. For methodological reasons, the 8th edition of the TNM staging system for NSCLC makes no official changes to the N descriptor. However, there is evidence that different subforms of N1 disease are associated with different prognoses, and it is now recommended that clinicians record the number of affected lymph nodes and nodal stations for further analyses. In this investigation we sought to determine whether patients with different levels and types of N1 lymph node involvement had significantly different 5-year survival rates. We retrospectively identified 90 patients with NSCLC (61 men, 29 women), who were treated between 2008 and 2012 and found to have pathologic N1 lymph node involvement and tumor sizes corresponding to T1 or T2. All patients were treated in curative intent with surgical lung resection and systematic mediastinal and hilar lymph node dissection. The overall 5-year survival rate was 56.3%. In the univariate analysis, lower tumor stage and tumor histology other than large-cell carcinoma were significantly associated with better long-term survival. Patients with solitary lymph node metastases also had longer disease-free survival than those with multiple nodal metastases. In the multivariate analysis, large-cell carcinoma and Union for International Cancer Control (UICC) stage IIB were independently associated with worse survival, while pneumonectomy, compared to lobar or sublobar resection, was independently associated with better survival. Although we did not observe significant prognostic differences between N1 subcategories within our patient population, other analyses may yield different results. Therefore, these data highlight the need for large, well-designed multicenter studies to confirm the clinical significance of N1 subcategories.

Integrative gene expression profiling reveals that dysregulated triple microRNAs confer paclitaxel resistance in non-small cell lung cancer via co-targeting MAPT.

BackgroundPaclitaxel has shown significant anti-tumor activity against non-small cell lung cancer (NSCLC); however, resistance to paclitaxel frequently occurs and represents a significant clinical problem and its underlying molecular mechanism remains elusive.MethodsLong-term treatment of culture cell with paclitaxel was carried out to mimic the development of acquired drug resistance in NSCLC. Cell proliferation and clonogenic assay and apoptosis evaluation were carried out to determine the efficacy of paclitaxel on NSCLC cells. Western blot analyses were performed to determine the expression and activation of proteins. Apoptosis enzyme-linked immunosorbent assay was used to quantify cytoplasmic histone-associated DNA fragments. Microarray analyses were applied to explore both mRNA and miRNA expression profiles in NSCLC cells followed by integrative analysis. qRT-PCR was carried out to verify the differentially expressed mRNAs and miRNAs.ResultsThe expression of 652 genes was shown to be changed at least 2-fold in paclitaxel-resistant NSCLC (H460_TaxR) cells with 511 upregulated and 141 downregulated as compared with that in parental H460 cells. The differentially expressed genes were functionally enriched in regulating the cell proliferation, cell death, and response to endogenous stimulus, and clustered in pathways such as cancer and signaling by the G protein-coupled receptor (GPCR). Moreover, 43 miRNAs were shown to be differentially expressed in H460_TaxR cells with 15 upregulated and 28 downregulated as compared with parental H460 cells. A total of 289 pairs of miRNA-potential target gene were revealed in H460_TaxR cells by bioinformatics analysis. Furthermore, integrative analysis of miRNAs and gene expression profiles revealed that dysregulated miR-362-3p, miR-766-3p, and miR-6507-3p might confer paclitaxel resistance in NSCLC via targeting MAPT simultaneously.ConclusionOur findings suggested that specific manipulation of MAPT-targeting miRNAs may be a novel strategy to overcome paclitaxel resistance in patients with NSCLC especially large-cell lung carcinoma.

Protein expression of close homologue of L1 (CHL1) is a marker for overall survival in non-small cell lung cancer (NSCLC).

The cell adhesion molecule close homologue of L1 (CHL1) is a potential tumour suppressor and was recently detected in non-small cell lung cancer (NSCLC) specimens. The expression pattern, prognostic, and functional role of CHL1 in NSCLCs is unknown. We evaluated the protein expression of CHL1 by immunohistochemistry in 2161 NSCLC patients based on a tissue microarray. The results were correlated with clinical, histopathological, and patient survival data (Chi square test, t test, and log-rank test, respectively). A multivariate analysis (Cox regression) was performed to validate its impact on patients' survival. CHL1 was expressed in NSCLC patients and was significantly overexpressed in lung adenocarcinomas and squamous cell carcinomas compared to neuroendocrine and large cell carcinomas of the lung (p < 0.001). CHL1 expression was associated with the T stage in adenocarcinomas (p = 0.011) and with metastatic lymph node status and UICC stage in squamous cell carcinomas (p = 0.034 and p = 0.035, respectively). Increased CHL1 expression was associated with improved survival in univariate (p = 0.031) and multivariate analyses (odds ratio 0.797, 95% confidence interval 0.677-0.939, p = 0.007). The prognostic significance of CHL1 makes it a potential prognostic and therapeutic target and underlines its role as a tumour suppressor. Further validation studies and functional analyses are needed to investigate its potential role in tumourigenesis and dissemination.

Abstract 3047: Connective tissue growth factor: Novel therapeutic target for non-small cell lung cancer

Abstract Lung cancer is the leading cause of cancer deaths worldwide. Non-small cell lung cancer (NSCLC), which accounts for 85% of all lung cancer cases, includes adenocarcinoma, squamous cell lung carcinoma and large-cell lung carcinoma. Despite recent advances in targeted therapies for adenocarcinoma, they are effective in only a small percentage of patients while there are no targeted therapies available for squamous cell lung cancer. Therefore, it is critical to identify novel intervention strategies against NSCLC. In the present study, we have identified connective tissue growth factor (CTGF) as a possible therapeutic target for NSCLC. CTGF is an extracellular-matrix (ECM) associated protein, which is involved in cellular processes like cell adhesion, migration, proliferation, differentiation, survival, apoptosis and senescence. CTGF has been associated with both oncogenic as well as tumor suppressive activities in context of different cancer types. The function of CTGF in lung cancer pathogenesis remains controversial. We performed immunohistochemistry on tissue microarray containing 60 cases, which clearly suggested that CTGF expression increases in lung carcinoma. More specifically, among other subtypes, squamous cell lung cancer cases expressed more CTGF compared to their matched normal tissues. To determine the functional role of CTGF in NSCLC, we employed both genetic and pharmacologic approaches to inhibit CTGF activity. Our results clearly demonstrated that CTGF inhibition by both lentiviral knockdown as well as monoclonal antibody against CTGF (FG-3019) caused marked inhibition in cell proliferation, colony formation and xenograft growth in immunodeficient mice. We have also found that CTGF knockdown causes cell cycle arrest and apoptosis. Furthermore, we tested the efficacy of FG-3019 along with drugs like cisplatin and gemcitabine in patient-derived xenograft (PDX) model of squamous cell lung cancer. Our results demonstrated that CTGF inhibition enhances the effects of gemcitabine chemotherapy in PDX model of squamous cell lung carcinoma. These results clearly suggested that CTGF has a profound effect on lung tumorigenesis, as its inhibition not only causes attenuation in tumor growth but also potentiates the chemotherapeutic sensitivity of commonly used drugs for NSCLC. We have also elucidated that tumor hypoxia and Yap signaling are responsible for CTGF up-regulation in lung carcinoma. Overall, our study underscores the significance of CTGF inhibition in non-small cell lung carcinoma and offers the potential use of FG-3019 for its treatment. Citation Format: Priyanka Chaudhary, Marie-Liesse Labat, Guangfang Wang, Mark Onaitis. Connective tissue growth factor: Novel therapeutic target for non-small cell lung cancer [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 3047.

Abstract 2021: Role of MMP1-PAR-1 crosstalk in the pro-tumorigenic senescent fibroblasts in large cell carcinoma of the lung

Abstract Tumor associated fibroblasts (TAFs) are key effector cells of cancer progression. Intriguingly, senescent TAFs have been reported in a growing list of agressive cancer subtypes including large cell carcinoma (LCC) of the lung. We previously reported that LCC cells induce fibroblast senescence in indirect co-cultures with transwells, revealing that paracrine signaling must be involved. A follow-up study identified MMP1 as an important regulatory protein in fibroblast senescence, since knocking-down MMP1 in LCC cells was sufficient to abrogate the induction of fibroblast senescence in co-cultures as well as the growth and invasion enhancement elicited by the conditioned medium of fibroblasts in LCC cells. Here we examined the potential role of the protease activated protein 1 (PAR-1) in fibroblast senescence by MMP1, since PAR-1 can be activated by MMP1 and has been reported in the stroma of lung cancer patients. We found that PAR-1 expression was down-regulated in fibroblasts that become senescent upon co-culture with LCC cells. In addition, forcing a reduced PAR-1 expression in fibroblasts by siRNA increased their expression of markers of the senescence associated secretory phenotype (SASP) during co-culture with LCC cells, whereas the percentage of senescence-associated beta-galactosidase positive fibroblasts remained largely unaffected. These results suggest that fibroblast senescence elicited by MMP1 secreted by LCC cells is mediated by processes other than PAR-1 activation and/or overexpression in fibroblasts. Citation Format: Marta Gabasa, Rafael Ikemori, Evette Radisky, Noemí Reguart, Derek Radisky, Jordi Alcaraz. Role of MMP1-PAR-1 crosstalk in the pro-tumorigenic senescent fibroblasts in large cell carcinoma of the lung [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 2021.

A study on the utility of ultrasound guided fnac in lung lesions

Introduction: Primary confirmation of Lung lesions by Ultrasound guided FNAC. It is useful for to differentiate Infective, Benign and Malignant lesions of lung. AIM: To study and confirm Cytological features of different types of lung lesions. 2. To assess the overall utility of FNAC in lung Pathology. Methods: Present study done over a period of two years, from Feb 2017- Jan 2019. During the above period 57 cases are studied, in the Department of pathology, Govt Medical College, Kadapa, AP. The above cases were categorised based on cytological features obtained by FNAC. Materials required for FNAC – 10 ml sterile disposable syringe with 25-22G needles. Suitable fixative is Isopropyl Alcohol. Alcohol fixed slides taken up for H&E staining. Results: Ultrasound guided FNAC of the Lung were performed on 57 cases. Out of 57 cases 48 (84.21%) cases were Primary neoplastic lesions, which are Squamous cell carcinoma, followed by large cell carcinoma 7 (12.28%) cases, Adeno carcinoma 2 (3.50%) cases.

Insulinoma-associated protein 1 is a sensitive and specific marker for lung neuroendocrine tumors in cytologic and surgical specimens

Insulinoma-associated protein 1 (INSM1) is an immunohistochemical marker for neuroendocrine differentiation with potentially superior sensitivity and specificity. INSM1 performance in pulmonary cytology cell block material (CB) has not been well established, and large series demonstrating its performance have been few. Typical and atypical carcinoid, small cell lung carcinoma, and large cell neuroendocrine carcinoma, squamous cell carcinoma, and adenocarcinoma CBs and 563 surgical specimens comprising 17 typical carcinoid, 14 atypical carcinoid, 8 small cell lung carcinoma, 10 large cell neuroendocrine carcinoma, 58 squamous cell carcinoma, 415 adenocarcinoma, and 17 large cell carcinoma cases and 24 other tumor types were immunostained with INSM1, CD56, synaptophysin, and chromogranin A. The INSM1 sensitivity, specificity, positive predictive value, and negative predictive value were 92.3%, 100%, 78.9%, and 99% in the CBs and 89.8%, 98.1%, 81.5%, and 99% in the surgical specimens, respectively, with 86.2% concordance. The sensitivity, specificity, positive predictive value, and negative predictive value for the other neuroendocrine markers were 97.4%, 93.3%, 97.4%, and 93.3% in the CBs and 93.9%, 93.6%, 58.2%, and 99.4% in the surgical specimens for CD56; 89.7%, 100%, 100%, and 75% in the CBs and 93.4%, 91.2%, 50.5%, and 99.4% in the surgical specimens for synaptophysin; 66.7%, 100%, 100%, and 53.6% in the CBs and 75.5%, 98.6%, 84.1%, and 97.7% in the surgical specimens for chromogranin A, respectively. Finally, INSM1, together with CD56, maximized the sensitivity to 100% with 93.3% specificity in the CBs. The results from our study have further established the high sensitivity and specificity of INSM1 in the largest pulmonary cytologic and surgical cohorts to date. INSM1 either matched or outperformed the performance of existing neuroendocrine markers, and its combination with CD56 appeared to maximize test performance.

Analysis of the clinical characteristics of 8 081 primary lung cancer

Objective: To characterize the clinical epidemiological features of primary lung cancer patients based on massive clinical data. Methods: The demographic and histological information of 8 081 primary lung cancer patients who were initially identified from 1 January 2012 to 31 December 2013 in 16 hospitals from 6 provinces were retrospectively analyzed to determine the characteristics of different histological subtypes among different gender, age-group and birth cohort. Results: Among the 8 081 lung cancer patients, 5 422 (67.10%) were male and 2 659 (32.90%) were female, the average age was (60.96±10.14) years. The most common histological subtypes of lung cancer successively were adenocarcinoma (ADC), squamous cell carcinoma (SCC), small cell carcinoma and large cell carcinoma, and the proportions of these subtypes were 53.13%, 24.51%, 14.59% and 0.66%, respectively, which collectively accounted for 92.89%. The current highest smoking rate was observed in SCC patients, which was 62.45%, while lowest in ADC, which was 29.68%. The incidence of lung cancer in male was significantly higher that that of female, with a sex ratio of 2.04∶1. The highest sex ratio was observed in SCC, which was 9.14∶1, while lowest in ADC, which was 1.14∶1. The distribution of histological subtypes in male lung cancer patients was consistent with the general situation.While among the female lung cancer patients, the proportion of ADC was the highest (75.42%), followed by SCC (10.08%), squamous cell carcinoma (7.34%) and large cell carcinoma (0.39%). Analyzed by the birth cohort, the proportion of ADC gradually increased with the age, while the reduced tendency was observed in SCC (P<0.000 1). Conclusion: Adenocarcinoma is the most important histological subtype of lung cancer, and the distribution characteristics of histological subtypes of lung cancer differs among genders, age and birth cohort.

Constitutional abnormality of nuclear membrane proteins in small cell lung carcinoma.

Nuclear membrane proteins reportedly play important roles in maintaining nuclear structures and coordinating cell activities. Studying profiles of nuclear membrane proteins may help us evaluate the biological and/or clinical nature of malignant tumors. Using immunohistochemistry with antibodies for emerin, lamin A/C, lamin B, and LAP2, we examined 105 lung cancer tissues from 33 small cell lung carcinomas (SCLCs) and 72 non-SCLCs (34 adenocarcinomas, 30 squamous cell carcinomas, and 8 large cell carcinomas). Emerin had negative or local/weak positivity in 79% of SCLCs and 1% of non-SCLCs, and lamin A/C had similar positivity in 91% of SCLCs and 3% of non-SCLCs. LAP2's expression was similar between SCLCs and non-SCLCs. RT-PCR analyses for these four nuclear membrane proteins over 7 cell lines showed that mRNA of emerin and lamin A/C were distinctly downregulated in the SCLC cell lines, supporting the immunohistochemical results. In conclusion, we suggest that downregulation of the nuclear membrane proteins emerin and lamin A/C is characteristic of SCLC cells, and this constitutional abnormality of the nuclear membrane may be related to the biological and/or clinical nature of SCLC. In addition, knowing the nuclear protein profile in SCLC cells may contribute to our understanding of nuclear fragility known as the crush artifact in pulmonary biopsy specimens.

Identification of key modules and hub genes for small-cell lung carcinoma and large-cell neuroendocrine lung carcinoma by weighted gene co-expression network analysis of clinical tissue-proteomes.

Small-cell lung carcinoma (SCLC) and large-cell neuroendocrine lung carcinoma (LCNEC) are high-grade lung neuroendocrine tumors (NET). However, comparative protein expression within SCLC and LCNEC remains unclear. Here, protein expression profiles were obtained via mass spectrometry-based proteomic analysis. Weighted gene co-expression network analysis (WGCNA) identified co-expressed modules and hub genes. Of 34 identified modules, six were significant and selected for protein–protein interaction (PPI) network analysis and pathway enrichment. Within the six modules, the activation of cellular processes and complexes, such as alternative mRNA splicing, translation initiation, nucleosome remodeling and deacetylase (NuRD) complex, SWItch/Sucrose Non-Fermentable (SWI/SNF) superfamily-type complex, chromatin remodeling pathway, and mRNA metabolic processes, were significant to SCLC. Modules enriched in processes, including signal recognition particle (SRP)-dependent co-translational protein targeting to membrane, nuclear-transcribed mRNA catabolic process of nonsense-mediated decay (NMD), and cellular macromolecule catabolic process, were characteristically activated in LCNEC. Novel high-degree hub genes were identified for each module. Master and upstream regulators were predicted via causal network analysis. This study provides an understanding of the molecular differences in tumorigenesis and malignancy between SCLC and LCNEC and may help identify potential therapeutic targets.

Computer-aided Quantification of Pulmonary Fibrosis in Patients with Lung Cancer: Relationship to Disease-free Survival.

Background Interstitial lung abnormalities (ILAs) are present at CT in about 10% of individuals undergoing lung cancer screening. The relationship between histologic findings of ILAs and their influence on patient prognosis remains unknown. Purpose To evaluate the percentage of ILAs at preoperative CT, as measured by radiologists and a computer-aided detection (CAD) software, as a predictor of disease-free survival in patients with lung cancer. Materials and Methods This retrospective study evaluated 217 consecutive patients who underwent complete resection of lung cancer from April 2010 to December 2015. Two radiologists, blinded to the patients' clinical information, scored percentage fibrosis extent and determined whether ILAs and the usual interstitial pneumonia (UIP) pattern were present. They assessed ILA progression at follow-up CT. Two pathologists determined the presence of an UIP pattern. Percentage fibrosis extent was also measured by using CAD. Binary logistic regression analysis was performed to determine whether the CAD percentage fibrosis extent was associated with ILA at CT. Multivariable Cox regression analysis was used to evaluate the significance of CAD percentage fibrosis extent as a predictor of prognosis. Results The radiologists classified 47 patients with ILAs and 24 patients with a UIP pattern at chest CT. The pathologists detected a UIP pattern in 25 patients. CT abnormalities showed progression over a 2-year period in all patients with histologic evidence of UIP. After adjustment for age, sex, and smoking index, the CAD percentage fibrosis extent was independently associated with ILA (odds ratio: 3.1; 95% confidence interval [CI]: 2.1, 4.7; P < .001). After adjustment for age, forced expiratory volume in 1 second (percentage predicted) radiologist-assessed percentage fibrosis extent, and pathologic stage, CAD percentage fibrosis extent was independently associated with worse disease-free survival (hazard ratio: 1.3; 95% CI: 1.1, 1.6; P < .001). Conclusion Greater computer-aided detection percentage fibrosis extent at preoperative CT independently predicted worse disease-free survival in patients with lung cancer. © RSNA, 2019 Online supplemental material is available for this article. See also the editorial by Goo in this issue.