Cases Of Non-small Cell Lung Carcinoma Research Articles

Prognostic Significance of N-Glycolyl GM3 Ganglioside Expression in Non-Small Cell Lung Carcinoma Patients: New Evidences.

The prognostic role of N-glycolyl GM3 ganglioside (NeuGcGM3) expression in non-small cell lung carcinoma (NSCLC) still remains controversial. In this study, the NeuGcGM3 expression was reevaluated using an increased number of NSCLC cases and the 14F7 Mab (a highly specific IgG1 raised against NeuGcGM3). An immunohistochemical score integrating the percentage of 14F7-positive cells and the intensity of reaction was applied to reassess the relationship between NeuGcGM3 expression, some clinicopathological features, and the overall survival (OS) of NSCLC patients. The double and the triple expression of NeuGcGM3 with the epidermal growth factor receptor (EGFR) and/or its ligand, the epidermal growth factor (EGF), were also evaluated. NeuGcGM3 expression correlates with both S-Phase fraction (p = 0.006) and proliferation index (p = 0.000). Additionally, NeuGcGM3 expression was associated with a poor OS of patients in both univariate (p = 0.020) and multivariate (p = 0.010) analysis. Moreover, the double and/or the triple positivity of tumors to NeuGcGM3, EGFR, and/or EGF permitted us to identify phenotypes of NSCLC with a more aggressive biological behavior. Our results are in agreement with the negative prognostic significance of NeuGcGM3 expression in NSCLC patients. However, standardization of techniques to determine the expression of NeuGcGM3 in NSCLC as well as the implementation of a universal scoring system is recommended.

Significance of DEK overexpression for the prognostic evaluation of non-small cell lung carcinoma.

In the present study, we explored the role of DEK expression for the prognostic evaluation of non-small cell lung carcinoma (NSCLC). DEK protein and mRNA expression levels were detected in NSCLC cells and fresh tissue samples of NSCLC paired with adjacent non-tumor tissues, respectively. NSCLC cases (n=196) meeting strict follow-up criteria were selected for immunohistochemical staining of DEK protein. Correlations between DEK expression and clinicopathological features of the NSCLC cases were evaluated using Chi-square tests. Survival rates were calculated using the Kaplan-Meier method, and the relationship between prognostic factors and patient overall survival was analyzed using Cox proportional hazard analysis. Based on the results, the levels of DEK protein and mRNA were significantly upregulated in 6 fresh tissue samples of NSCLC. Immunohistochemical analysis showed that the DEK expression rate was significantly higher in the NSCLC samples compared with either the adjacent non-tumor tissues or normal lung tissues. DEK expression was correlated with poor differentiation and late pathological stage of NSCLC. DEK expression was also correlated with low disease-free survival and overall survival rates. In the early-stage group, disease-free and overall survival rates of patients with DEK expression were significantly lower than those of patients without DEK expression. Further analysis using a Cox proportional hazard regression model revealed that DEK expression emerged as a significant independent hazard factor for the overall survival rate of patients with NSCLC. Consequently, DEK plays an important role in the progression of NSCLC. DEK may potentially be used as an independent biomarker for the prognostic evaluation of NSCLC.

Targeting stemness is an effective strategy to control EML4-ALK+ non-small cell lung cancer cells.

The fusion between anaplastic lymphoma kinase (ALK) and echinoderm microtubule-associated protein-like 4 (EML4) is a causative factor in a unique subset of patients with non-small cell lung carcinoma (NSCLC). Although the inhibitor crizotinib, as it blocks the kinase activity of the resulting EML4-ALK fusion protein, displays remarkable initial responses, a fraction of NSCLC cases eventually become resistant to crizotinib by acquiring mutations in the ALK domain or activating bypass pathways via EGFR, KIT, or KRAS. Cancer stem cell (CSC) theory provides a plausible explanation for acquisition of tumorigenesis and resistance. However, the question as to whether EML4-ALK-driven tumorigenesis is linked with the stem-like property and whether the stemness is an effective target in controlling EML4-ALK+ NSCLC including crizotinib-resistant NSCLC cells has not been addressed. Here, we report that stem-like properties stem from ALK activity in EML4-ALK+ NSCLC cells. Notably, treatment with rapamycin, a CSC targeting agent, attenuates stem-like phenotypes of the EML4-ALK+ cells, which increased capability of tumor formation and higher expression of stemness-associated molecules such as ALDH, NANOG, and OCT4. Importantly, combinational treatment with rapamycin and crizotinib leads to synergistic anti-tumor effects on EML4-ALK+ NSCLC cells as well as on those resistant to crizotinib. Thus, we provide a proof of principle that targeting stemness would be a novel strategy to control intractable EML4-ALK+ NSCLC.

Comparison of immunohistochemistry and mRNA in situ hybridization in detecting thyroid transcription factor-1 expression in non-small cell lung carcinomas tissue

Thyroid transcription factor-1 (TTF-1) reportedly possesses oncogenic and suppressive roles within the same tumor type and may play a dual function in the progression of lung cancer. Immunohistochemistry (IHC) and mRNA in situ hybridization (ISH) are commonly used methods for detecting protein or mRNA expression. The present study compared the concordance rate of the two methods in the evaluation of thyroid transcription factor-1 (TTF-1) expression in non-small cell lung carcinoma (NSCLC) using tissue microarray-based IHC and mRNA ISH. TTF-1 protein and mRNA expression levels were examined in 196 cases of NSCLC. The IHC and mRNA ISH agreement was 91.3% (179/196), and near-perfect agreement was observed between the two methods (κ-coefficient, 0.848). There was no significant difference between IHC and mRNA ISH, as analyzed by the McNemar-Bowker test (P=0.219). The present findings proved that IHC is comparable to mRNA ISH for evaluating TTF-1 expression in NSCLC. These two methods can be used to detect TTF-1 expression in future studies.

Abstract 3181: EGFR and KRAS activation generate discrete inflammatory responses within the lung tumor microenvironment

Abstract Despite decades of research, non-small cell lung carcinoma (NSCLC) remains one of the world's deadliest cancers. Approximately half of NSCLC cases are classified as lung adenocarcinomas (ADCA), in which KRAS, EGFR, and TP53 mutations are the predominant genetic drivers. Although patients with EGFR mutations are initially responsive to targeted therapies, drug resistance uniformly occurs within one year. KRAS mutant ADCA have proven even more intractable, as scant progress has been made in the development of therapeutics for this patient population. Recent breakthroughs in the field of immunotherapy have offered new hope for the management of NSCLC, but the clinical success of immunomodulatory agents will depend on a strong foundational knowledge of the immune cells that comprise the ADCA microenvironment. Activating mutations in KRAS and EGFR induce the aberrant function of numerous signaling pathways that may influence the host immune response in different ways. However, the discrete impact of KRAS and EGFR mutations on the immune composition of ADCA remains largely undefined. To address this question, we profiled the tumor microenvironments (TME) of four mouse models of ADCA: adenoviral Cre-infected KrasLSL-G12D and KrasLSL-G12D;Trp53Fl/Fl mice, as well as TetO-EgfrL858R;Ccsp-rtTA and TetO-EgfrL858R;T790M;Ccsp-rtTA mice. Lung specimens from tumor-bearing and control animals (n ≥ 5 per cohort) were harvested at multiple time points and subjected to histological assessment, gene expression analysis by qRT-PCR, and flow cytometric immunophenotyping, in which 23 markers were used to identify 12 unique cell types. The Kras subtype exhibited robust inflammation and expansion of both the lymphoid and myeloid cell compartments. In particular, significant increases were observed in the number of CD8+ T cells (3-fold), B cells (2.5-fold), and macrophages (16-fold). Although loss of Trp53 promoted tumor growth and malignancy, it had little effect on cytokine expression or immune cell recruitment into the Kras TME. In contrast, while EgfrL858R mutations produced similar total levels of inflammation, leukocyte recruitment into the Egfr TME was largely restricted to macrophage expansion (increased 8-fold). Introduction of the T790M resistance mutation to Egfr-mutant animals further amplified the macrophage dominant signal (14-fold). Expression of the cytokine IL-6 was correspondingly elevated in the Egfr but not Kras subtypes, a finding replicated in EGFR-transfected primary human endobronchial cells. Most notably, CD8+ cytotoxic T cell numbers were unaltered at all time points examined in the Egfr animals, a finding which raises concerns about the efficacy of most currently developed immune checkpoint blockers in EGFR patients. In summary, KRAS and EGFR mutations give rise to distinct immune responses that may require oncogene-specific immunotherapy regimens. Citation Format: Stephanie E. Busch, Mark L. Hanke, Kyoung Hee Kim, Julia Kargl, A. McGarry Houghton. EGFR and KRAS activation generate discrete inflammatory responses within the lung tumor microenvironment. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3181. doi:10.1158/1538-7445.AM2015-3181

The role of immunohistochemical analysis in the evaluation of EML4-ALK gene rearrangement in lung cancer.

Among the mutations described in non-small cell lung carcinoma is a rearrangement resulting from an inversion within chromosome 2p leading to the formation of a fusion gene, echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase (EML4-ALK). Fluorescence in situ hybridization (FISH) is the gold standard for the detection of ALK gene rearrangements. However, molecular methods are not readily available in all pathology laboratories. Immunohistochemistry (IHC) using an antibody directed against the EML4-ALK fusion protein provides a widely available alternative method of detection. We assessed whether IHC is a comparable and cost-effective alternative to FISH analysis for the detection of ALK gene rearrangements. A total of 110 non-small cell lung carcinoma cases (63 surgical/biopsy and 47 cytology specimens), previously tested for ALK gene rearrangements by FISH [7 (6.4%) positive for the rearrangement], were probed for the EML4-ALK fusion protein using a monoclonal EML4-ALK antibody, clone 5A4. Cells were considered to stain positive for ALK if >5% of cells showed cytoplasmic staining of at least grade 1 intensity (scale: 0 to 3). A cost analysis was performed using ALK IHC as a screening test. The sensitivity and specificity of the EML4-ALK IHC stain compared with ALK FISH analysis were 100% and 96%, respectively. All 7 FISH-positive cases stained positive by IHC, whereas 4 FISH-negative cases demonstrated positive staining. One of the 4 FISH-negative, IHC-positive cases harbored an EML4-ALK rearrangement by RT-PCR yielding 3 false-positive results overall. The κ agreement between IHC and FISH methods is 0.76 (substantial/excellent). The potential savings of implementing the ALK IHC as a screening method would be $10,418.21. Sensitivity of the EML4-ALK IHC stain is excellent (100%) but due to its suboptimal specificity, IHC cannot reliably supplant FISH analysis for the detection of ALK gene rearrangements. IHC shows promise as a screening tool to prevent unnecessary costly FISH analysis.

Expression of hypoxia inducible factor-1α, hypoxia inducible factor-2α and vascular endothelial growth factor in metastatic lung cancer and clinical significance

Objective To discuss the expression patterns of hypoxia inducible factor(HIF)-1α, HIF- 2α and vascular endothelial growth factor(VEGF)in metastatic lung cancer tissues. Methods The immunohistochemical streptavid-in- peroxidase(SP) method was applied to detect the expression of HIF-1α, HIF- 2α and VEGF in 93 cases of surgicall resected metastatic lung cancer, and 57 cases of primary non- small- cell lung carcinoma(NSCLC)as blank control group. Results HIF-1α, HIF-2α and VEGF proteins were expressed in 72 cases of metastatic lung cancer and 58 cases of primary NSCLC with the positive rate being 77.4%, 62.3% and 74.2% respectively(P 0.05). Conclusion HIF-1α, HIF-2α and VEGF expression in lung metastases is increased, and HIF-1α expression was positively correlated with VEGF expression. Key words: Metastatic lung cancer; Hypoxia inducible factor-1α; Hypoxia inducible factor-2α; Vascular endothelial growth factor

Accuracy of classifying poorly differentiated non-small cell lung carcinoma biopsies with commonly used lung carcinoma markers.

Immunohistochemical (IHC) staining is an important adjunct to the classification of non-small cell lung carcinoma (NSCLC). Several studies have used tissue microarrays derived from resection specimens to evaluate the accuracy of IHC staining for classifying NSCLC, but few have used actual biopsies of poorly differentiated carcinomas, and the question of how often biopsy IHC in such tumors leads to the correct classification has received little attention. We identified 40 cases of NSCLC that, on biopsy, could not be subclassified by morphology and that had subsequent resection specimens. TTF-1, napsin, p63/p40, and CK5 or a subset thereof were used for IHC classification. Of the 40 cases classified by IHC on biopsy, 33 (82%) had no change in diagnosis after resection. Of the remaining 7 cases, 3 were classified as NSCLC--not otherwise specified on biopsy and subclassified as either adenocarcinoma or squamous cell carcinoma (SCC) on the surgical specimen. One adenocarcinoma biopsy was reclassified as pleomorphic carcinoma. Two SCCs were changed to adenosquamous carcinoma, and 1 SCC was changed to large cell lung carcinoma. Only 1 antibody pair (2%) was discordant between biopsy, and almost all reclassifications were done based on morphologic features rather than change in IHC pattern. We conclude that IHC staining allows accurate subclassification of poorly differentiated NSCLCs on small lung biopsies in most cases, but there is still a substantial "miss" rate (here, 18%). Surgical resection specimens allow further subclassification, mainly due to architectural features not present in the biopsies.

Performance of a multiplexed dual analyte immunoassay for the early detection of non-small cell lung cancer.

Objectives“PAULA’s” test (Protein Assays Utilizing Lung cancer Analytes) is a novel multiplex immunoassay blood test that incorporates both tumor antigens and autoantibodies to determine the risk that lung cancer (LC) is present in individuals from a high-risk population. The test’s performance characteristics were evaluated in a study using 380 retrospective clinical serum samples.MethodsPAULA’s test is performed on the Luminex xMAP technology platform, and detects a panel of 3 tumor antigens (CEA, CA-125, and CYFRA 21–1) and 1 autoantibody marker (NY-ESO-1). A training set (n = 230) consisting of 115 confirmed diagnoses of non-small cell lung carcinoma (NSCLC) cases and 115 age- and smoking history-matched controls was used to develop the LC predictive model. Data from an independent matched validation set (n = 150) was then used to evaluate the model developed, and determine the ability of the test to distinguish NSCLC cases from controls.ResultsThe 4-biomarker panel was able to discriminate NSCLC cases from controls with 74% sensitivity, 80% specificity, and 0.81 AUC in the training set and with 77% sensitivity, 80% specificity, and 0.85 AUC in the independent validation set. The use of NY-ESO-1 autoantibodies substantially increased the overall sensitivity of NSCLC detection as compared to the 3 tumor markers alone. Overall, the multiplexed 4-biomarker panel assay demonstrated comparable performance to a previously employed 8-biomarker non-multiplexed assay.ConclusionsThese studies confirm the value of using a mixed panel of tumor antigens and autoantibodies in the early detection of NSCLC in high-risk individuals. The results demonstrate that the performance of PAULA’s test makes it suitable for use as an aid to determine which high-risk patients need to be directed to appropriate noninvasive diagnostic follow-up testing, especially low-dose CT (LDCT).

Correlation of Cytomorphology and Molecular Findings in EGFR+, KRAS+, and ALK+ Lung Carcinomas

There is increasing emphasis on the subclassification of non-small cell lung carcinomas (NSCLCs) and molecular features to guide treatment. Histologic studies have suggested some morphologic features predominating in tumors. Our aim was to determine if mutated cases had distinct cytomorphology. A retrospective study was designed to retrieve all cytopathology cases of NSCLC that had mutation studies for EGFR or KRAS or fluorescence in situ hybridization studies for ALK between 2007 and 2012. All slides from available mutation-positive cases were reviewed, and cytomorphologic features were correlated to mutation status. Of the cases with molecular testing, 62 (39%) of 160 were positive for mutation, including 39 (31%) positive for KRAS, 20 (14%) for EGFR, and 4 (3%) for ALK (one case positive for both EGFR and ALK). More of ALK+ and KRAS+ cases had a diagnosis of NSCLC-favor adenocarcinoma or NSCLC not otherwise specified than did EGFR+ cases (25; 51% vs 5%). Eosinophilic granular cytoplasm was seen in more ALK and KRAS cases than in EGFR cases (100; 32% vs 6%). Cytologic features of ALK+ and KRAS+ tumors included more nuclear pleomorphism, necrosis, and a less vacuolated cytoplasm than did EGFR+ tumors, which may explain the less definitive subclassification in ALK+ and KRAS+ tumors.

Metastasis of primary lung carcinoma to the breast: a systematic review of the literature

BackgroundThe purpose of this systematic review was to summarize previously published case reports of primary lung carcinoma metastasis to the breast to assess common clinical and pathologic features and management strategies. Materials and methodsCase reports describing breast metastasis of primary lung carcinoma were systematically evaluated in MEDLINE and EMBASE. ResultsThirty-one reported cases of non-small-cell lung carcinoma (NSCLC) metastasized to the breast were identified, along with eight cases of small-cell lung carcinoma. Sixty-seven percent of reported NSCLC metastases to the breast were detected metachronously with the primary lung abnormality, whereas 80% of small-cell lung carcinoma breast metastases appeared synchronously. Thyroid transcription factor 1 was found to be expressed in 58% of total NSCLC breast metastases, including 83% of those of adenocarcinoma origin. Therapeutic strategies among NSCLC cases varied widely, and only 36% of NSCLC breast metastasis patients were administered chemotherapy. Additional sites of metastasis in these cases are summarized as well. ConclusionsIt is recommended to include metastatic lung cancer in the differential diagnosis of patients presenting with a breast abnormality in the context of a suspected lung cancer. Thyroid transcription factor 1 expression should be examined in these cases. The metachronous versus synchronous nature of lung carcinoma metastasis to the breast has consequences for both detection of the primary and secondary lesions and patient outlook. Clinical correlation is vital to effective management of the care of patients harboring these atypical secondary lesions.

A Single-Tube Multiplexed Assay for Detecting ALK, ROS1, and RET Fusions in Lung Cancer

Approximately 7% of non-small cell lung carcinomas (NSCLCs) harbor oncogenic fusions involving ALK, ROS1, and RET. Although tumors harboring ALK fusions are highly sensitive to crizotinib, emerging preclinical and clinical data demonstrate that patients with ROS1 or RET fusions may also benefit from inhibitors targeting these kinases. Using a transcript-based method, we designed a combination of 3' overexpression and fusion-specific detection strategies to detect ALK, ROS1 and RET fusion transcripts in NSCLC tumors. We validated the assay in 295 NSCLC specimens and showed that the assay is highly sensitive and specific. ALK results were 100% concordant with fluorescence in situ hybridization (FISH) (n = 52) and 97.8% concordant with IHC (n = 179) [sensitivity, 96.8% (95% CI 91.0%-98.9%); specificity, 98.8% (95% CI 93.6%-99.8%)]. For ROS1 and RET, we also observed 100% concordance with FISH (n = 46 and n = 15, respectively). We identified seven ROS1 and 14 RET fusion-positive tumors and confirmed the fusion status by RT-PCR and FISH. One RET fusion involved a novel partner, cutlike homeobox 1 gene (CUX1), yielding an in-frame CUX1-RET fusion. ROS1 and RET fusions were significantly enriched in tumors without KRAS/EGFR/ALK alterations. ALK/ROS1/RET/EGFR/KRAS alterations were mutually exclusive. As a single-tube assay, this test shows promise as a more practical and cost-effective screening modality for detecting rare but targetable fusions in NSCLC.

Pharmacogenetics of DNA repair gene polymorphisms in non-small-cell lung carcinoma patients on platinum-based chemotherapy

Individual differences in chemosensitivity and clinical outcome of non-small-cell lung carcinoma (NSCLC) patients can be influenced by host-inherited factors. We investigated the impact of XRCC1 Arg194Trp, XRCC1 Arg280His, XRCC1 Arg399Gln, XPD Arg156Arg, XPD Asp312Asn, XPD Asp711Asp, and XPD Lys751Gln gene polymorphisms on treatment efficacy in 375 NSCLC patients on platinum-based chemotherapy. We also examined progression-free survival and overall survival. The gene polymorphisms were analyzed by duplex PCR. The patients with XRCC1 399A/A had a significantly better response to chemotherapy. Individuals with XPD 711 Asp and XPD 312 Asn alleles responded poorly to chemotherapy when compared with the wide-type genotype. The adjusted hazard ratio (HR) in the Cox regression model was calculated. The XRCC1 399A/A polymorphism was associated with better progression free survival and overall survival of NSCLC patients (HR=0.61 and 0.55). On the other hand, the XPD 711 Asp allele was associated with poorer progression free survival and overall survival compared to the C/C genotype, with HRs of 1.89 and 1.90. The XPD 312 Asn allele was found to be associated with non-significantly reduced survival of NSCLC patients (HR = 1.73). In conclusion, we found the polymorphisms of XRCC1 and XPD to be related to the efficacy of platinum-based chemotherapy in NSCLC patients. This information should aid in therapeutic decisions for individualized therapy in NSCLC cases.

Treatment of mid-late stage NSCLC using sodium cantharidinate/vitamin B6/GP regimen in clinic.

The present study observed the clinical outcomes of the sodium cantharidinate/vitamin B6/GP regimen in the treatment of middle-late stage Non-small-cell lung carcinoma (NSCLC). Eighty-six cases of NSCLC were selected and randomized into two groups. Forty-five cases in the treatment group were subject to 30 ml cantharidinate/vitamin B6+ GP regimen, for four 21-day cycles. Forty-one cases in the control group were subject to regular GP regimen, for four 21-day cycles. The effectiveness rate was 57.8% in the treatment group and 36.6% in the control group, suggesting statistical difference (P < 0.05); life quality in the treatment group was significantly better than that in the control group (P < 0.05); the occurrence rate of the toxic/adverse effects were significantly lower in the treatment group compared with that in the control group (P < 0.05). Sodium cantharidinate/vitamin B6/GP regimen had fair effectiveness and synergistically improved the clinical outcomes. It lowered the toxic/adverse effects and its application is worth further investigation and promotion.

Validation of a Histology-Independent Prognostic Gene Signature for Early-Stage, Non–Small-Cell Lung Cancer Including Stage IA Patients

Patients with early-stage non-small-cell lung carcinoma (NSCLC) may benefit from treatments based on more accurate prognosis. A 15-gene prognostic classifier for NSCLC was identified from mRNA expression profiling of tumor samples from the NCIC CTG JBR.10 trial. In this study, we assessed its value in an independent set of cases. Expression profiling was performed on RNA from frozen, resected tumor tissues corresponding to 181 stage I and II NSCLC cases collected at University Health Network (UHN181). Kaplan-Meier methodology was used to estimate 5-year overall survival probabilities, and the prognostic effect of the classifier was assessed using the log-rank test. A Cox proportional hazards model evaluated the signature's effect adjusting for clinical prognostic factors. Expression data of the 15-gene classifier stratified UHN181 cases into high- and low-risk subgroups with significantly different overall survival (hazard ratio [HR] = 1.92; 95% confidence interval [CI], 1.15-3.23; p = 0.012). In a subgroup analysis, this classifier predicted survival in 127 stage I patients (HR = 2.17; 95% CI, 1.12-4.20; p = 0.018) and the smaller subgroup of 48 stage IA patients (HR = 5.61; 95% CI, 1.19-26.45; p = 0.014). The signature was prognostic for both adenocarcinoma and squamous cell carcinoma cases (HR = 1.76, p = 0.058; HR = 4.19, p = 0.045, respectively). The prognostic accuracy of a 15-gene classifier was validated in an independent cohort of 181 early-stage NSCLC samples including stage IA cases and in different NSCLC histologic subtypes.

Prediction effects of vimentin expression in non-small cell lung cancer metastases

Objective To examine vimentin expression in non-small cell lung carcinomas (NSCLC) by using immunohistochemistry to evaluate potential correlations between vimentin expression and the metastase of the patients.Methods 295 cases of NSCLC including 164 cases of squamous cell carcinomas (SCC),108 cases of adenocarcinomas (AC) and 23 cases of other NSCLC carcinomas were examined by using immunohistochemistry.Results Vimentin was indeed detected in 145 cases (49.2％).It was principally present in cells at the tumour/stroma interface.Vimentin expression was significantly associated with the differentiation status of carcinomas.The follow-up of patients further demonstrated that an extensive expression of vimentin (＞50％ of tumour cells) was associated with the occurrence of metastases.Conclusion Our data demonstrate that vimentin expression is a frequent event in NSCLC and that its expression can be associated with the occurrence of metastases positively. Key words: Non small cell lung cancer; Vimentin; Differentiation; Metastases

Impact of HER2 and PTEN Simultaneous Deregulation in Non-small Cell Lung Carcinoma: Correlation with Biological Behavior

HER2/neu overexpression due to gene amplification is an important factor in breast cancer, modifying the sensitivity to anti-HER2 monoclonal antibody therapy. The clinical significance of HER2 expression in non small cell lung carcinoma (NSCLC) is currently under evaluation. The tumor suppressor gene PTEN negatively regulates the HER2/PI3K/Akt signalling pathway. The purpose of this study was to evaluate the role of simultaneous alteration in HER2 and PTEN protein expression in relation to biological behaviour of NSCLCs. Protein expression was determined by immunohistochemistry in sixty-one (n=61) NSCLC cases along with CISH for HER2 gene analysis and detection of chromosome 17 aneuploidy. Patients were followed-up for a period of 34 to 41 months after surgery. HER2 overexpression (2+/3+ score) was detected in 17 (27.9%) patients while loss of PTEN expression was observed in 24 (39.3%) cases, low expression in 29 (47.6%) and overexpression in 8 (13.1%). Simultaneous HER2 overexpression and PTEN low/loss of expression were correlated with metastasis (71.4% vs 36.2% p=0.03) . Analysis in the subgroup of 22 patients of pTNM stage III with lymph node status N1 or N2 revealed that there was a relationship between the number of positive regional lymph node groups and simultaneous deregulation of the two genes (p=0.04). Multivariate analysis determined that HER2 overexpression was associated with an increasing risk of developing metastases (OR: 4.3; 95%CI: 1.2-15.9; p: 0.03) while PTEN overexpression was associated with lower risk (OR: 0.1; 95%CI: 0.1, 1.0; p: 0.05). Simultaneous HER2/PTEN deregulation is a significant genetic event that leads to a more aggressive phenotype of NSCLC.

Expression of miR-150 and miR-3940-5p is reduced in non-small cell lung carcinoma and correlates with clinicopathological features

The present study investigated the expression of miR-150 and miR-3940-5p in non-small cell lung carcinoma(NSCLC) and its relationship with clinicopathologic features. Samples included tumor, tumor-adjacent and normal lung parenchyma tissues from 90 NSCLC patients and 17 cases of embryonic lung cDNA. The expression levels of miR-150, miR-18b-5p, miR-643 and miR-3940-5p were detected by real‑time PCR; p53, EGFR, Kras and Ki-67 expression in tumor tissues was determined by immunohistochemistry. p53 mRNA expression levels in NSCLC were examined by SYBR-Green real-time PCR. The relationship between the four miRNAs and clinicopathologic features of 90 cases was analyzed. The expression of miR-150 and miR-3940-5p was significantly downregulated in tumor tissues and embryonic lung tissues compared to normal lung tissues. The expression of miR-150 and miR-3940-5p in tumor tissues was also lower than that in the matched tumor-adjacent tissues. miR-150 was downregulated preferentially in subgroups of patients with a tumor diameter more than or equal to 3cm, in smokers and in stageIII and IV tumors. Specifically, miR-150 and miR-3940-5p expression was decreased in nuclear cell proliferation antigen Ki-67-positive NSCLC cases. miR-150 and miR-3940-5p were found to be significantly downregulated in p53 IHC-positive NSCLC cases and were negatively correlated with p53 mRNA. Reduced miR-150 and miR-3940-5p expression in tumor tissues and embryonic lung tissues suggests that these miRs may be involved in the tumorigenesis or de-differentiation of NSCLC. Due to this associaton with the Ki-67 proliferation index in NSCLC, downregulation of miR-150 and miR-3940-5p may contribute to tumor growth and proliferation. miR-150 and miR-3940-5p may affect p53 expression through a direct or indirect pathway.

ROS1 Receptor Tyrosine Kinase, a Druggable Target, is Frequently Overexpressed in Non-Small Cell Lung Carcinomas Via Genetic and Epigenetic Mechanisms

Microarray analyses have revealed significantly elevated expression of the proto-oncogene ROS1 receptor tyrosine kinase in 20-30% of non-small cell lung carcinomas (NSCLC). Selective and potent ROS1 kinase inhibitors have recently been developed and oncogenic rearrangement of ROS1 in NSCLC identified. We performed immunohistochemical evaluation of expression of ROS1 kinase and its downstream molecules in 399 NSCLC cases. ROS1 expression in primary and recurring lesions of 92 recurrent NSCLC cases was additionally analyzed. To elucidate mechanism of expression, two ROS1-nonexpressing NSCLC cell lines (Calu6 and H358) and fresh frozen tissues from 28 consecutive NSCLC patients were examined for ROS1 promoter methylation status and ROS1 expression. Overall expression rate of ROS1 was 22% (19% for adenocarcinomas and 25% for nonadenocarcinomas) in NSCLC. ROS1 expression was a worse prognostic factor for overall survival in adenocarcinomas of stage I NSCLC. In recurred NSCLC, ROS1 expression was significantly higher in recurring tumors (38%) than primary tumors (19%). Two NSCLC cell lines showed increased ROS1 expression after treatment with 5-aza-2'deoxycytidine and/or trichostatin A. Among the 14 adenocarcinomas examined, two (14%) showed more than twice the level of ROS1 expression in tumor tissue than was observed in matched normal tissue and statistically significant differences in the ROS1 promoter methylation level. A subset of NSCLC revealed overexpression of ROS1 receptor tyrosine kinase, possibly in relation to epigenetic changes. ROS1 expression was an independent prognostic factor for overall survival in adenocarcinomas of stage I NSCLC. Further studies are needed to validate our results.

Anaplastic lymphoma kinase gene rearrangement in non-small-cell lung cancer in a Brazilian population

Lung cancer is the leading cause of cancer deaths worldwide (1). Non-small-cell lung carcinoma (NSCLC), which represents approximately 80% of all lung cancer cases, is frequently diagnosed at advanced stages of the disease and is associated with short survival time. Although the prognosis of this disease remains poor, significant advances in the genetics and treatment of NSCLC have been made in recent years. In 2007, Soda et al. (2) identified the transforming EML4-ALK fusion gene in 6.7% of NSCLC patients. This fusion transcript, which presents potent oncogenic activity both in vitro and in vivo (2,3), is formed by the translocation of the echinoderm microtubule-associated protein-like 4 (EML4) gene (2p21) and anaplastic lymphoma kinase (ALK) gene (2p23). Subsequent studies have shown that between two and eight percent of all NSCLC cases may harbor ALK rearrangements, which are almost always mutually exclusive with EGFR and KRAS mutations. Moreover, these patients are frequently younger, non- or light smokers and have adenocarcinomas (2),. Recently, crizotinib, an orally available small-molecule inhibitor of the ALK tyrosine kinase, was approved for the treatment of ALK-rearranged NSCLC patients, with successful results (24-29). The two most widely used methods for the diagnosis of ALK-rearranged NSCLC are fluorescence in situ hybridization (FISH) and immunohistochemistry. Because there is no information about the prevalence and clinical characteristics of ALK-rearranged NSCLC in Latin America in the literature, the aim of this study was to evaluate NSCLC with ALK rearrangement in Brazilian patients.