Chang Li Wang

Value of the metastatic lymph node ratio for predicting the prognosis of non-small-cell lung cancer patients.

BackgroundThe aim of this study was to investigate the relation between the metastatic lymph node ratio (LNR) and the prognosis of non-small-cell lung cancer (NSCLC).MethodsA total of 301 patients with N1 or N2 NSCLC who underwent complete pulmonary resection were analyzed retrospectively. The correlations between the LNR and clinical and pathologic data were analyzed using χ2 test analysis. The prognostic value of the LNR was calculated by univariate Kaplan–Meier survival analysis and multivariate Cox proportional hazard model analysis. The risk groups were classified by a combination of the LNR and pN stage.ResultsThe LNR was correlated with age, smoking status, pathologic type, subcarinal lymph node, clinical staging, N stage (Pxa0<xa00.05), and the number of positive lymph nodes and positive lymph node stations (Pxa0<xa00.0001). In the univariate analysis, the LNR played an important role in predicting overall survival (OS) (Pxa0<xa00.0001) and disease-free survival (Pxa0<xa00.0001) by Kaplan–Meier survival analysis. In the multivariate analysis, high LNR (>18%) was an independent poor prognostic factor for OS [hazard ratio (HR) 2.5034, 95% confidence interval (CI) 1.6096–3.8933, Pxa0<xa00.0001] and DFS (HR 1.9023, 95% CI 1.2465–2.9031, Pxa0=xa00.0031). Stratification into high-, medium-, and low-risk groups—based on high-risk factors (LNRxa0>xa018%, N2) intermediate-risk factors (LNRxa0>xa018%, N1 or LNRxa0<xa018%, N2), and low-risk factors (LNRxa0<xa018%, N1)—could efficiently predicted outcomes (Pxa0<xa00.0001) of patients with lymph node-positive NSCLC.ConclusionsThe combination of the LNR and pN status provides a valuable help with prognosis. However, these results must be evaluated further in a large prospective randomized clinical trial.

Combined Erlotinib and Cetuximab overcome the acquired resistance to epidermal growth factor receptors tyrosine kinase inhibitor in non-small-cell lung cancer

PurposeNon-small-cell lung cancer (NSCLC) cells with somatic mutations in epidermal growth factor receptors (EGFR) are initially susceptible to tyrosine kinase inhibitor (TKI); however, eventually resistance to TKI is developed in these cells, which leads to the failure of treatment. The most common mechanism of this acquired drug resistance is development of a secondary T790M mutation in EGFR. In this study, we investigated the effects of the combination of Erlotinib and Cetuximab on T790M and L858R mutation lung cancer cells lines (H1975), in the primary NSCLC cells with the T790M mutation and TKI-resistant EGFR mutations human tumor xenograft model (H1975).MethodsThe effects of these two agents on cell proliferation, apoptosis, and EGFR-dependent signaling were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, annexin V staining, and Western blotting. Sensitivity of EGFR inhibitors was detected in the primary tumor cell suspension and human tumor xenograft model (H1975).ResultsCompared with single-agent treatment, the combination of Cetuximab and Erlotinib increased apoptosis of EGFR TKI–resistant NSCLC cells (H1975), resulting in more pronounced growth inhibition on cell proliferation and significant inhibition of EGFR-dependent signaling.ConclusionsThese data suggest that treatment with a combination of Erlotinib and Cetuximab overcomes T790M-mediated drug resistance.

Osteopontin Combined With CD44v6, a Novel Prognostic Biomarker in Non-Small Cell Lung Cancer Undergoing Curative Resection

BACKGROUNDnOsteopontin (OPN) is identified as one of the leading genes that promote the metastasis of malignant tumor through binding to CD44v6 and integrin. The purpose of the current study was to assess the prognostic significance of OPN and CD44v6 in patients with non-small cell lung cancer (NSCLC).nnnMETHODSnTissue microarray was used to detect the expression of OPN and CD44v6 in 159 NSCLC patients undergoing complete pulmonary resection in our hospital between 2003 and 2006. The correlations among OPN, CD44v6, and clinicopathologic data were analyzed using χ(2) testing analysis. The prognostic values of OPN and CD44v6 were evaluated by univariate Kaplan-Meier survival analysis and multivariate Cox proportional hazard model analysis.nnnRESULTSnOPN and CD44v6 were both independent predictors for overall survival and disease-free survival. When OPN and CD44v6 were considered together, the predictive range was extended and the sensitivity was improved, especially for those patients with stage I NSCLC. The 6-year overall survival and disease-free survival rates in OPN+ or CD44v6+ patients were 49.1% and 39.6%, respectively, which were significantly lower than those of OPN-/CD44v6- patients (64.4% and 47.7%, respectively), and were higher than those of OPN+/CD44v6+ patients (16.4% and 14.8%, respectively). Stratification into OPN+/CD44v6+, OPN+ or CD44v6+, or OPN-/CD44v6- groups, based on the expression OPN and CD44v6, could efficiently predicted outcomes (p < 0.001) of NSCLC patients.nnnCONCLUSIONSnThe combination of OPN and CD44v6 is a valuable independent predictor of tumor recurrence and survival in NSCLC patients.

CCR1 knockdown suppresses human non-small cell lung cancer cell invasion

PurposeCC chemokine receptor 1 (CCR1) plays a critical role in the recruitment of leukocytes to the site of inflammation. Tumor invasion and metastasis share many similarities with leukocyte trafficking, which is critically regulated by chemokines and their receptors. In this study, we aimed to assess the role of CCR1 in non-small cell lung cancer (NSCLC).MethodsCCR1 expression was determined by Western blotting in two human NSCLC clones (95C and 95D) with different metastatic potential. We silenced CCR1 expression through microRNA-mediated RNA interference, and examined the invasiveness and proliferation of CCR1-silenced NSCLC cell through Matrigel assay and MTT assay. Matrix metalloproteinases (MMPs) activity was determined by gelatin zymography.ResultsWe found that expression of CCR1 was correlated with the aggressive phenotype of the NSCLC cells. CCR1 knockdown significantly suppressed the invasiveness of NSCLC cells, but had only a minor effect on cell proliferation. Moreover, we demonstrated that CCR1 knockdown significantly reduced the expression level of matrix metalloproteinase-9.ConclusionsThese findings suggest that CCR1 contributes to NSCLC cell migration by stimulating cell invasion, independent of cell proliferation. CCR1 might be a new target for NSCLC therapy.

Osteopontin knockdown suppresses non-small cell lung cancer cell invasion and metastasis

Background Osteopontin (OPN) was identified as one of the leading genes that promote the metastasis of malignant tumor. However, the mechanism by which OPN mediates metastasis in non‐small cell lung cancer (NSCLC) remains unknown. The aim of the study is to investigate the biological significance and the related molecular mechanism of OPN expression in lung cancer cell line. Methods Lentiviral‐mediated RNA interference was applied to inhibit OPN expression in metastatic human NSCLC cell line (A549). The invasion, proliferation, and metastasis were evaluated OPN‐silenced in A549 cells in vitro and in vivo. The related mechanism was further investigated. Results Interestingly, OPN knockdown significantly suppressed the invasiveness of A549 cells, but had only a minor effect on the cellular migration and proliferation. Moreover, we demonstrated that OPN knockdown significantly reduced the levels of matrix metalloproteinase (MMP)‐2 and urokinase plasminogen activator (uPA), and led to an obvious inhibition of both in vitro invasion and in vivo lung metastasis of A549 cells (P <0.001). Conclusions Our data demonstrate that OPN contributes to A549 cell metastasis by stimulating cell invasion, independent of cellular migration and proliferation. OPN could be a new treatment target of NSCLC.

Do tumor cavitation and sex in resected stage I non-small-cell lung cancer correlate with prognosis?

BackgroundMultiple factors have been reported as affecting the prognosis, and they affect the therapeutic outcomes of stage I non-small-cell lung cancer (NSCLC) patients. Most studies focus on patients receiving combined-modality therapy, whereas there are few studies that focus on patients undergoing surgery alone. The aim of this study was to identify risk factors for disease relapse and unfavorable prognosis in stage I NSCLC patients treated with surgery alone.MethodsA total of 315 stage I NSCLC patients who were treated with surgery alone as the definitive therapy were identified. Risk factors for disease relapse and unfavorable prognosis were estimated by univariate and multivariate analyses.ResultsSex, tumor pathologic stage, and cavitating lung cancer were identified as independent risk factors for relapse and overall survival using the multivariate analysis. Sex, tumor pathologic stage, and cavitating lung cancer were identified as independent risk factors for early relapse, and sex and cavitating lung cancer were independent risk factors for late relapse.ConclusionTumor cavitation, pathologic stage IB, and being male are predictors of poor outcome for patients with stage I NSCLC who undergo resection.

Osteopontin-expressing macrophages in non-small cell lung cancer predict survival

BACKGROUNDnTumor-associated macrophages (TAMs) are a major component of leukocyte infiltration in the tumor microenvironment. Osteopontin is related to tumor metastasis and proliferation. Osteopontin is expressed not only by tumor cells but also by TAMs. The purpose of the current study was to assess the prognostic significance of osteopontin expressed by TAMs (TOPN) in patients with non-small cell lung cancer.nnnMETHODSnTissue microarray was used to detect the expression of TOPN, TAMs, and microvascular density in 159 patients with non-small cell lung cancer undergoing complete pulmonary resection in our hospital between 2003 and 2006. The correlations between TOPN, TAMs, and clinicopathologic data were analyzed with χ(2) tests. Quantitation of TAMs or TOPN and microvascular density analyses was performed using Bonferroni correction and the Students t test. The prognostic value of TOPN was evaluated by univariate Kaplan-Meier survival analysis and multivariate Cox proportional hazard model analysis.nnnRESULTSnIn the recurrence and metastasis group, microvascular density was higher than that in the control group (14.4 ± 1.06 versus 8.9 ± 1.02; pxa0= 0.0002). In the TOPN-positive group, microvascular density was increased compared with that in the TOPN-negative group (14.3 ± 1.37 versus 10.7 ± 0.91; pxa0= 0.0273). Osteopontin expressed by TAMs was an independent predictor for overall survival (pxa0= 0.017) and disease-free survival (p < 0.001), especially for stage I non-small cell lung cancer. The 6-year overall and disease-free survival rates in TOPN-positive patients were 22.64% and 16.98%, respectively, which were significantly lower than those of TOPN-negative patients (50.00% and 39.62%, respectively).nnnCONCLUSIONSnOsteopontin expressed by TAMs is a valuable independent predictor of tumor recurrence and survival in patients with non-small cell lung cancer.

Claudin-1 correlates with poor prognosis in lung adenocarcinoma

This study was conducted to investigate the clinical significance of claudin‐1 (CLDN1) expression in patients with lung adenocarcinoma.

Expressions of CLDN1 and insulin-like growth factor 2 are associated with poor prognosis in stage N2 non-small cell lung cancer.

Background Patients with single station mediastinal lymph node (N2) non‐small cell lung cancer (NSCLC) have a better prognosis than those with multilevel N2. The molecular factors which are involved in disease progression remain largely unknown. The purpose of this study was to investigate gene expression differences between single station and multilevel N2 NSCLC and to identify the crucial molecular factors which are associated with progress and prognosis of stage N2 NSCLC. Methods Gene expression analysis was performed using Agilent 4×44K Whole Human Genome Oligo Microarray on 10 freshfrozen lymph node tissue samples from single station N2 and paired multilevel N2 NSCLC patients. Real‐time reverse transcription (RT)‐PCR was used to validate the differential expression of 14 genes selected by cDNA microarray of which four were confirmed. Immunohistochemical staining for these validated genes was performed on formalin‐fixed, paraffin‐embedded tissue samples from 130 cases of stage N2 NSCLC arranged in a high‐density tissue microarray. Results We identified a 14 gene expression signature by comparative analysis of gene expression. Expression of these genes strongly differed between single station and multilevel N2 NSCLC. Four genes (ADAM28, MUC4, CLDN1, and IGF2) correlated with the results of microarray and real‐time RT‐PCR analysis for the gene‐expression data in samples from 56 NSCLC patients. Immunohistochemical staining for these genes in samples from 130 cases of stage N2 NSCLC demonstrated the expression of IGF2 and CLDN1 was negatively correlated with overall survival of stage N2 NSCLC. Conclusions Our results suggest that the expression of CLDN1 and IGF2 indicate a poor prognosis in stage N2 NSCLC. Further, CLDN1 and IGF2 may provide potential targeting opportunities in future therapies.

Global gene expression differentiating pure bronchioloalveolar carcinoma from adenocarcinoma with bronchioloalveolar carcinoma features

OBJECTIVESnPure bronchioloalveolar carcinoma (BAC) is considered the early stage of lung adenocarcinoma, and is even regarded as lung adenocarcinoma in situ. This study was designed to investigate the differences in the gene expression of pure BAC and that of adenocarcinoma with bronchioloalveolar features and explore the mechanism of BAC progression to adenocarcinoma with bronchioloalveolar featuresnnnMETHODSnTotal RNA was extracted from 16 tissue specimens. Expression analysis was carried out using Agilent 4 × 44 k arrays. Gene ontology analysis was used to define pathways altered in bronchioloalveolar progression. Differentially expressed candidate genes were validated using quantitative real-time PCR. The statistical analysis was carried out according to the methods of the paired t-test.nnnRESULTSnAdenocarcinoma with bronchioloalveolar features demonstrated an increased expression of 23 genes and reduced expression of 20 genes compared with BAC. These genes were considered candidate marker genes for tumour progression and metastasis. Genes overexpressed in adenocarcinoma with bronchioloalveolar features included fibroblast growth factor receptor 1, and CLDN18 (claudin 18), whereas those overexpressed in BAC included ataxia telangiectasia and Rad3 related (ataxia telangiectasia mutated and Rad3-related), and activating transcription factor 2. Mitogen-activated protein kinase (MAPK) pathway seemed dysregulated in adenocarcinoma with bronchioloalveolar features compared with pure BAC.nnnCONCLUSIONSnMicroarray-based expression profiling revealed interesting novel candidate genes in BAC and adenocarcinoma with bronchioloalveolar features. The MAPK pathway seemed dysregulated in adenocarcinoma with bronchioloalveolar features compared with the pure BAC pathway, which is worthy of being explored because it could partially explain the mechanism of the progression of BAC to adenocarcinoma with bronchioloalveolar features.