Value In Non-small Cell Lung Cancer Research Articles

Expression of adiponectin in non-small cell lung cancer and its relationship with MMP-9 and angiogenesis

To explore the expression of adiponectin and its relationship with matrix metalloproteinase (MMP)-9 and angiogenesis in non-small cell lung cancer (NSCLC) tissue. A total of 87 samples of NSCLC and 10 samples of normal lung tissue were collected for detection of adiponectin and MMP-9 expression by immunohistochemical method. Anti-CD34 monoclonal antibody against endothelial marker was used to evaluate microvessel density (MVD). The positive rate for adiponectin in NSCLC (49.43%) was significantly lower than that in normal lung tissues (90.00%) while for MMP-9 in NSCLC (59.77%) was significantly higher than that in normal lung tissues (10.00%, P<0.05); MVD in NSCLC (14.58 ± 0.67) was higher than that in normal lung tissues [(0.80 ± 0.12), P<0.05]. In NSCLC, the positive rates of adiponectin and MMP-9 were varied in groups with different lymph node metastasis, tumor size and clinical stage (P<0.05). Meanwhile, MVD value in NSCLC with or without lymph node metastasis was significantly different. There was a negative correlation between adiponectin level and CD34 (MVD) or MMP-9 level (r=-0.22 or r=-0.55, respectively, P<0.05). On the contrary, MMP-9 was positively correlated with MVD (r=0.21, P<0.05). The decreased expression of adiponectin might be related to the development of NSCLC through negative regulation of MMP-9 expression. .

High levels of Phosphatase and Tensin Homolog Expression Predict Favorable Prognosis in Patients with Non-small Cell Lung Cancer.

The prognostic role of phosphatase and tensin homolog (PTEN) in non-small cell lung cancer (NSCLC) has been controversial. In this study, levels of PTEN expression were investigated in NSCLC patients and their prognostic value in NSCLC was assessed. PTEN expression in tumor tissues from 68 NSCLC patients was analyzed using immunohistochemistry and confocal microscopy. Survival analysis was performed using the log-rank test and Cox proportional hazards regression analysis. NSCLC patients classified as expressers of high levels of PTEN (n=46) had better prognoses than those classified as expressers of low levels (mean survival 17.1 vs. 12.9months, log rank P=0.038). In patients with adenocarcinoma (AC), high PTEN expression (n=9) was associated with significantly longer survival than low PTEN expression (mean survival 23.50 vs. 15.54months, log rank P=0.043). High levels of PTEN expression resulted in 43% reduction in risk for all NSCLC patients (HR 0.57, 95% CI 0.33-0.98, P=0.041). PTEN expression and clinical stage remained significantly associated with survival after adjustment for age, sex, and tumor type (HR 0.56, 95% CI 0.32-0.99; P=0.048; HR 0.54, 95% CI 0.36-0.97; P=0.045). No significant difference in continuous PTEN expression levels was observed among groups with different clinical or pathological characteristics (P>0.17). When levels of PTEN expression were binarized using the optimal cut-point, higher levels of PTEN expression were observed in patients with T1/T2 than in those with T3/T4 (80 and 58%, respectively, P=0.049) and in patients with AC than in those with squamous-cell carcinoma (78 and 58%, respectively, P=0.08). No significant difference in binarized PTEN expression levels was found among groups with any other clinical/pathologic characteristic (P>0.28). Our results suggest that high levels of PTEN expression may be favorable prognostic markers in NSCLC patients.

Prognostic impact of COX-2 in non-small cell lung cancer: A comprehensive compartment-specific evaluation of tumor and stromal cell expression

Cyclooxygenase-2 (COX-2) is an enzyme that has been extensively investigated as a prognostic marker in cancer. In non-small cell lung cancer (NSCLC) previous results regarding the prognostic impact of COX-2 expression are inconsistent. Therefore we evaluated the association between transcript levels and overall survival in nine publicly available gene expression data sets (total n = 1337) and determined in situ compartment-specific tumor and stromal cell protein expression in two independent cohorts (n = 616). Gene expression did not show any correlation with clinical parameters or with overall survival. Protein expression in tumor and stromal cells did not correlate with any clinical parameter or with overall survival in one of the analyzed cohorts, while a significant association of high stromal expression with longer survival was observed in both univariate and multivariate analysis in the other cohort. Stromal expression of COX-2 has not been separately evaluated in NSCLC previously and may be a subject of further investigation, whereas the presented findings from this comprehensive compartment specific evaluation clearly reject the hypothesis of COX-2 tumor cell expression having a prognostic value in NSCLC.

WE‐E‐17A‐05: Complementary Prognostic Value of CT and 18F‐FDG PET Non‐Small Cell Lung Cancer Tumor Heterogeneity Features Quantified Through Texture Analysis

Purpose:Previous studies have shown that CT or 18F‐FDG PET intratumor heterogeneity features computed using texture analysis may have prognostic value in Non‐Small Cell Lung Cancer (NSCLC), but have been mostly investigated separately. The purpose of this study was to evaluate the potential added value with respect to prognosis regarding the combination of non‐enhanced CT and 18F‐FDG PET heterogeneity textural features on primary NSCLC tumors.Methods:One hundred patients with non‐metastatic NSCLC (stage I–III), treated with surgery and/or (chemo)radiotherapy, that underwent staging 18F‐FDG PET/CT images, were retrospectively included. Morphological tumor volumes were semi‐automatically delineated on non‐enhanced CT using 3D SlicerTM. Metabolically active tumor volumes (MATV) were automatically delineated on PET using the Fuzzy Locally Adaptive Bayesian (FLAB) method. Intratumoral tissue density and FDG uptake heterogeneities were quantified using texture parameters calculated from co‐occurrence, difference, and run‐length matrices. In addition to these textural features, first order histogram‐derived metrics were computed on the whole morphological CT tumor volume, as well as on sub‐volumes corresponding to fine, medium or coarse textures determined through various levels of LoG‐filtering. Association with survival regarding all extracted features was assessed using Cox regression for both univariate and multivariate analysis.Results:Several PET and CT heterogeneity features were prognostic factors of overall survival in the univariate analysis. CT histogram‐derived kurtosis and uniformity, as well as Low Grey‐level High Run Emphasis (LGHRE), and PET local entropy were independent prognostic factors. Combined with stage and MATV, they led to a powerful prognostic model (p&lt;0.0001), with median survival of 49 vs. 12.6 months and a hazard ratio of 3.5.Conclusion:Intratumoral heterogeneity quantified through textural features extracted from both CT and FDG PET images have complementary and independent prognostic value in NSCLC.

Decreased miRNA-148a is associated with lymph node metastasis and poor clinical outcomes and functions as a suppressor of tumor metastasis in non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death worldwide, and only 15% of lung cancer patients live more than 5 years. microRNAs (miRNAs) are endogenously expressed non-coding RNAs, and dysregulation of miRNAs is a common feature in human cancers including lung cancer. In this study, we describe the epigenetic regulation of miRNA-148a and its prognostic value in NSCLC. Due to hypermethylation of the miRNA‑148a encoding region, the expression levels of miRNA-148a were decreased in NSCLC tissues and cells. Decreased miRNA‑148a expression was associated with lymph node metastasis, advanced clinical stage and shortened disease-free survival and overall survival in NSCLC, and was an independent prognostic factor for overall survival in multivariate analysis. In vitro, overexpression of miRNA-148a significantly suppressed the migratory and invasive abilities of A549 and H1299 lung cancer cells. Enforced expression of miRNA-148a in lung cancer cell lines resulted in a significant reduction in the expression of DNMT1. This, in turn, led to a decrease in DNA methylation of the tumor-suppressor gene E-cadherin and induced an increase in the protein levels of E-cadherin. By understanding the function and molecular mechanism of miRNA-148a in NSCLC, miRNA-148a may have therapeutic potential to suppress lung cancer metastasis.

Mucin 1 (MUC1) expression in patients (pts) with early stage non-small cell lung cancer (NSCLC): Relationship between immunohistochemistry (IHC), tumor characteristics, and survival.

3011 Background: MUC1, a glycoprotein highly expressed in many malignancies, is being explored as an antigen for immunotherapy. How best to measure MUC1 expression as well as its prognostic value in NSCLC are still under discussion. Methods: TMAs were constructed using triplicate 1mm cores of FFPE tumour and stained with 214D4 (recognises protein core) and MA695 (recognises carbohydrate epitope) anti-MUC1 antibodies (abs). TMAs were assessed for polarisation, both cyto and mem staining intensity (scored 0-3) and proportion cells +ve (0-100%; scored 0–5), averaged for multiple cores. A composite score (intensity x cells +ve) was derived, ranging from 0–15 (3+ in &gt;75% cells). Results: TMAs from 521 pts were analysed: male 362 (69.5%); never smoking 35 (7%); adeno 259 (49.7%), squamous 180 (34.5%), large cell 39 (7.5%); nodal N0 340 (65.3%), N1 71 (13.6%), N2 107 (20.5%). Results of IHC staining intensity, proportion positive cells and depolarisation were very similar for the two abs. There was high concordance in the composite score for the abs (R2=0.71, p&lt;0.0001). Polarisation was discordant in 7.9% of cases. For 77 cases with paired primary/ N2 nodal tissue, mean 214D4 scores were 8.3 and 8.9 and MA695 10.6 and 9.9 respectively. Discordant staining in primary but not in node was seen in 5.2% and 10.4% with 214D4 and MA695 respectively. Increased expression as assessed by 214D4 (HR=1.26; 95% CI 1.014-1.565, p=0.04 log rank test) and MA695 (HR=1.20, 95% CI 1.042-1.605; p=0.02) was associated with improved survival. The prognostic value of depolarisation has yet to be analyzed. Conclusions: Over 93% of cases were MUC1 IHC positive. Composite scores for the 2 abs were highly correlated and depolarisation largely concordant. MUC1 expression was generally maintained in paired primary/nodal tumour. Increased expression of MUC1 was associated with improved survival however further investigation is needed to determine which abs best predict outcomes. [Table: see text]

Negative NKX2-1 (TTF-1) as Temporary Surrogate Marker for Treatment Selection During EGFR-Mutation Analysis in Patients with Non–Small-Cell Lung Cancer

In the past decade, major progress has been made toward personalized medical treatment of non-small-cell lung cancer (NSCLC) through the discovery of epithelial growth factor receptor (EGFR) mutations. However, mutation analysis takes extra time and additional costs in the diagnostic evaluation of lung cancer patients. It has been hypothesized that EGFR mutations are restricted to terminal respiratory unit -type adenocarcinoma expressing thyroid transcription factor-1 (official symbol NKX2-1) as determined by immunohistochemistry. The aim of the current study is to evaluate the potential of NKX2-1 immunohistochemistry as a prescreening test for EGFR mutation analysis. From 2004 to December 2010, 810 consecutive NSCLC tumor specimens were tested for EGFR mutations in a routine diagnostic procedure. Immunohistochemistry for NKX2-1 was performed (clone 8G7G3/1 [Dako]) and the results were compared with tumor EGFR-mutation status and clinicopathological characteristics. EGFR mutations were detected in 114 specimens (14%). NKX2-1 expression was present in 68%. In the cases with EGFR mutation, NKX2-1 staining was positive in 92%. NKX2-1 immunohistochemical (IHC) staining was significantly associated with the presence of EGFR mutations (p = 5.3×10). NKX2-1 increased the negative predictive value in NSCLC to more than 95%. In case of a negative NKX2-1 IHC staining, and only if clinically urgent, the high negative predictive value of more than 95% for EGFR mutations is a suitable temporary surrogate marker for the choice of starting with chemotherapy. In case of positive NKX2-1 IHC, the best strategy is to wait for the outcome of EGFR-mutation analysis and then choose the appropriate treatment.

Predictive impact of RRM1 protein expression on vinorelbine efficacy in NSCLC patients randomized in a chemotherapy phase III trial.

10617 Background: Platinum based doublets remain the cornerstone of treatment in non-small cell lung cancer (NSCLC) and often includes gemcitabine. A biomarker predicting sensitivity to this antimetabolite would represent a major step forward in the management of advanced disease. Ribonucleotide reductase subunit M1 (RRM1) has been of some value in NSCLC but evidence is not uniform. Accordingly, we explored the predictive role of RRM1 in patients with advanced NSCLC. Methods: 443 patients with advanced NSCLC were randomized in a phase III trial to regimen A (paclitaxel and cisplatin with gemcitabine) or regimen B (cisplatin and vinorelbine). Immunohistochemical evaluation of RRM1 was performed on mainly bioptic material and correlated to response rates, progression free survival (PFS) and overall survival (OS). Results: 261 (58.9%) patients had representative tissue samples for RRM1 evaluation. Disease control rate, PFS and OS were significantly improved in patients with RRM-negative tumors receiving regimen B as compared to patients with RRM-positive tumors (68.8% vs. 31.2%, P = 0.046 , 6.90 months (mths) (95% CI 5.83-7.96) vs. 3.93 mths (95% CI 3.11-4.76), P = 0.000 and 11.57 mths (95% CI 9.65-13.48) vs. 7.4 mths (95% CI 5.37-9.43), P = 0.002, respectively). However, this was not the case for patients receiving regimen A (67.6% vs. 32.4%, P = 0.366, 6.73 mths vs. 7.6 mths, P = 0.207, 11.07 mths vs. 12.37 mths, P = 0.103, respectively ). Together with histological subtype, RRM1-positivity emerged as the only other significant prognostic variable with a hazard ratio of 1.56 (95% CI 1.38-2.35, P = 0.033) in multivariate analysis for patients treated with regimen B. Conclusions: RRM1 protein expression was without predictive impact in patients treated with cisplatin, paclitaxel and gemcitabine. Surprisingly, the predictive power proved robust in the cisplatin and vinorelbine arm across classic endpoints confirmed by multivariate analysis. These findings may suggest that RRM1 is involved in vinorelbine sensitivity and warrant further research which will be presented shortly.

Highly expressed SLC35F2 in non-small cell lung cancer is associated with pathological staging

Homo sapiens solute carrier family 35 member F2 (SLC35F2) is highly homologous to the lung squamous cell cancer-related gene, LSCC3, which is highly expressed in lung squamous cell tumour tissues. However, the clinical implication of the SLC35F2 gene in tumour development and progression remains unclear. An affinity-purified polyclonal antibody raised against the human SLC35F2 peptide was used in the immunohistochemical analysis of a non-small cell lung cancer (NSCLC) tissue microarray of human NSCLC (n=129). SLC35F2 protein was also analysed with the same antibody using Western blotting. Total RNAs were extracted from tumour tissues (n=43) and from laser-dissected tumour cells (n=9). SLC35F2 gene expression was detected by fluorescent real-time quantitative PCR and compared to the expression in the corresponding adjacent normal lung tissues. It was found that both the SLC35F2 protein (by IHC analysis) and the SLS35F2 gene transcripts (by Q-PCR analysis) were expressed at significantly higher levels in the NSCLC tumour tissues than in the corresponding adjacent normal lung tissues (p<0.001 and =0.015, respectively). There was a significant correlation between the SLC35F2 transcript and pathological staging (r=0.219, p=0.029), although the correlation between SLC35F2 protein and the staging was not significant (r=0.175). SLC35F2 was highly expressed in NSCLC tissues and the levels of expression, in particular the levels of the SLC35F2 transcript, were associated with NSCLC pathological staging. SLC35F2 appears to have a significant prognostic value in NSCLC.

Class III β-Tubulin in Advanced NSCLC of Adenocarcinoma Subtype Predicts Superior Outcome in a Randomized Trial

Platinum-based doublets are the cornerstone of treatment in advanced non-small-cell lung cancer (NSCLC) and often include vinorelbine or taxanes. A predictive biomarker is greatly needed to select chemotherapy-sensitive patients for these microtubule-interfering agents. Class III β-tubulin (TUBB3) has been shown of value in NSCLC, but evidence is not uniform. Accordingly, we explored the predictive role of TUBB3 in advanced NSCLC. Four hundred forty-three patients with advanced NSCLC were enrolled in a phase III trial and randomized to vinorelbine- or paclitaxel-containing chemotherapy. Immunohistochemical evaluation of TUBB3 status was mainly done on bioptic material and correlated to response rates, progression-free survival (PFS), overall survival (OS), quality of life (QOL), and toxicity. Two hundred sixty-one (58.9%) patients had representative tissue samples for TUBB3 evaluation. Patients with TUBB3-negative adenocarcinomas had a significantly prolonged PFS and OS when compared with the opposite subgroup (7.87 vs. 6.83 months, P = 0.035 and 14.17 vs. 11.17 months, P = 0.018, respectively). Multivariate analyses revealed an HR of 1.55 (95% CI, 1.04-2.31, P = 0.032) for TUBB3-positive adenocarcinoma patients. TUBB3-negative adenocarcinoma patients showed a mean QOL decline of -18.25 points (95% CI, -4.28 to -32.22, P = 0.013) as compared with -3.86 (95% CI, -7.0 to 15.52, P = 0.5). TUBB3 was of predictive value in adenocarcinoma patients in the largest, randomized advanced NSCLC population published to date. It may be clinically useful in conjunction with other biomarkers, but QOL information should be recorded during validation, as prophylactic intervention may be needed in specific subgroups at risk of toxicity.

Preoperative lymphocyte count is an independent prognostic factor in node-negative non-small cell lung cancer

A number of prognostic factors have been reported in non-small cell lung cancer (NSCLC). Although lymph node metastasis is the most poorly predictive value in completely resected NSCLC, a significant number of patients have a fatal recurrence even in node-negative curative NSCLC. Recently inflammatory response has been shown as a predictive value in NSCLC. Neutrophils and lymphocytes play an important role in cancer immune response. In this study, we retrospectively examined the impact of preoperative peripheral neutrophil and lymphocyte counts on survival, and investigated the relationships of these factors to clinicopathological factors in node-negative NSCLC. A total 237 patients were evaluated. When the cut-off value of neutrophil count was 4500 mm −3 with a maximum log-rank statistical value, overall 5-year survival rates were 79.7% for the low-neutrophil-count group and 69.5% for the high-neutrophil-count group ( P = 0.04). When the cut-off value of lymphocyte count was 1900 mm −3 with a maximum log-rank statistical value, overall survival rates were 67.9% for the low-lymphocyte group and 87.7% for the high-lymphocyte group ( P < 0.001). High-neutrophil-counts were associated with tumor size ( P = 0.002) and pleural invasion ( P < 0.001). Low-lymphocyte-counts were correlated with vascular invasion ( P = 0.018) and recurrence of NSCLC ( P = 0.01). Multivariate analysis showed that the lymphocyte count was an independent prognostic factor (hazard ratio: 3.842; 95% confidence interval: 1.827–8.078; P < 0.001), but the neutrophil count was not ( P = 0.185). We conclude that a peripheral lymphocyte count, which is associated with vascular invasion, is an independent prognostic factor in node-negative NCSLC.

Phase II study of sunitinib as maintenance therapy in patients with locally advanced or metastatic non-small cell lung cancer

This open-label, phase II study evaluated the antitumor activity and safety of sunitinib monotherapy as maintenance treatment following first-line chemotherapy in patients with locally advanced or metastatic non-small cell lung cancer (NSCLC). Following treatment with standard doublet chemotherapy (paclitaxel and carboplatin), patients received oral sunitinib (starting dose 50mg/day) in 6-week cycles (Schedule 4/2: 4 weeks on treatment, 2 weeks off treatment) until disease progression, unacceptable toxicity or withdrawal of consent. The primary endpoint was probability of survival at 1 year ≥55%. Of 84 patients who received first-line chemotherapy, 66 (79%) received sunitinib maintenance therapy (median sunitinib cycles started: 2 [range 1–20]). Probability of survival at 1 year was 40.5% (95% confidence interval [CI]: 29.8, 51.0). Median overall survival was 10.4 months (95% CI: 8.0, 12.2). The objective response rate was 27.4% (95% CI: 18.2, 38.2). The most frequently reported all-causality adverse events of any grade during sunitinib maintenance therapy were fatigue/asthenia (55%), diarrhea (36%), and nausea (32%). These data suggest that maintenance therapy may have value in NSCLC, although the primary endpoint of the study was not met.

Standardization of Epidermal Growth Factor Receptor (EGFR) Measurement by Quantitative Immunofluorescence and Impact on Antibody-Based Mutation Detection in Non–Small Cell Lung Cancer

Challenges in measurement of epidermal growth factor receptor (EGFR) protein expression have led to conflicting data on its prognostic value and discontinuation of its use for prediction of response. Herein is described a quantitative standardized assay for EGFR and its use in a series of retrospective cohorts of patients with non-small cell lung cancer (NSCLC). The AQUA technology of quantitative immunofluorescence was used in conjunction with Western blot analysis to calculate the absolute concentration of EGFR in two independent NSCLC cohorts (170 from Yale New Haven Hospital and 335 from Sotiria and Patras Hospitals in Greece). EGFR and mutated EGFR were measured using D38B1 antibody and two mutation-specific antibodies. All patients positive or borderline for mutation-specific antibody were genotyped. A threshold for reproducible detection of EGFR was defined as 0.85 ng/μg total protein. EGFR expression demonstrated no prognostic value in either cohort. The mutation rate was 1.79% in the Yale cohort, and 1.52% in the Sotiria/Patras cohort, with no antibody detection-based false-positive cases. No mutations were detected for EGFR concentrations <1.46 ng/μg total protein. In summary, accurate measurement of EGFR still shows no prognostic value in NSCLC. In these two population-based cohorts, the antibody-based EGFR mutation rate was lower than has been frequently reported.

Abstract 160: Altered levels of circulating microRNAs in the plasma of non-small cell lung cancer patients: Potential as biomarker

Abstract The development of molecular markers for lung cancer could offset the deficiencies of currently available early detection and staging approaches. To this end, blood markers are particularly attractive since samples are obtained in a minimally invasive way at any stage of lung carcinogenesis. In this study we compared the levels of 10 microRNAs (miR-21, miR-31, miR-155, miR-147, miR-218, miR-377, miR-551b*, miR-569, miR-1283, and miR-1468) in the blood plasma of 27 non-small-cell lung cancer (NSCLC) patients and 38 healthy controls, using quantitative reverse transcription- polymerase chain reaction (qRT-PCR), to determine if these miRNAs might have a diagnostic value for NSCLC. The results of the area under the receiver operating characteristic curve (ROC), which measures the ability of the test to correctly classify those with and without the disease, were 0.98, 1.00, 0.8, and 0.98 for miR-21, miR-155, miR-377 and miR-1468, respectively, indicating that plasma levels of miR-21, miR-155, miR-377 and miR-1468 are powerful discriminators between patients and controls. At the optimum cutoff point for the cycle threshold (CT) value of the four miRNAs, the sensitivities of miR-21, miR-155, miR-377 and miR-1468 were 92.5%, 100%, 84%, and 100%, respectively; the corresponding values for the test specificity were 100%, 100%, 88%, and 100%, respectively. These results indicate the great potential of miR-21, miR-155, miR-377 and miR-1468 as diagnostic markers for NSCLC but need to be proved in prospective studies with larger samples. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 160. doi:10.1158/1538-7445.AM2011-160

Diagnostic and prognostic role of plasma levels of two forms of cytokeratin 18 in patients with non-small-cell lung cancer

Purpose Cytokeratin 18 (CK18) can be used as a serum biomarker for carcinoma cell death, whereas caspase-cleaved (ccCK18) fragments reflect tumour apoptosis. We explored the potential diagnostic and prognostic role of circulating CK18 and ccCK18 in patients with non-small-cell lung cancer (NSCLC) in comparison with Cyfra 21.1, a fragment of cytokeratin 19. Methods Subject cohorts consisted of 200 healthy blood donors (HBD), 113 patients with benign lung diseases (BLD) and 179 NSCLC cases. Plasma levels of ccCK18, total CK18 and Cyfra 21.1 were determined with ELISA assays. Results Plasma levels of ccCK18 and total CK18 were higher in the NSCLC group compared to the HBD and BLD cohorts ( p < 0.0001). Using a cut-off of 104 U/L for ccCK18 and 302 U/L for total CK18 (95% specificity in the HBD group) the diagnostic accuracy of both CK18 forms to distinguish between NSCLC and BLD cases was 56%, whereas it was 94% for Cyfra 21.1. Multivariate survival analysis showed that total CK18 was a stronger prognostic factor than both ccCK18 and Cyfra 21.1 (HR 0.64 for low versus high total CK18 levels, 95% confidence interval (CI) 0.50–0.82; p = 0.0004) in the entire NSCLC cohort and in 78 patients with locally advanced or metastatic disease treated with chemoradiotherapy or first-line chemotherapy (HR 0.70 95% CI 0.52–0.94; p = 0.018). Conclusions Cyfra 21.1 is a useful diagnostic biomarker for NSCLC. Total CK18 shows a promising potential as prognostic marker in NSCLC patients, independently of the therapeutical intervention. In contrast, ccCK18 was not of prognostic value in NSCLC, suggesting that tumour necrosis is of particular importance in this disease.

KRAS mutations (MTs) in non-small cell lung cancer (NSCLC) versus colorectal cancer (CRC): Implications for cetuximab therapy.

10529 Background: KRAS MTs have been associated with a diversity of biologic functions, and have both prognostic and predictive consequences in NSCLC and CRC. Little data exist comparing the specifics of KRAS MTs in the two cancers. In CRC, benefit from the cetuximab is mainly limited to patients whose tumors are KRAS wild-type (WT). However, recent studies show no such predictive value in NSCLC. We hypothesized that the unique molecular biology and tobacco carcinogen-related etiology of NSCLC contribute to a divergence in KRAS MT type and EGFR-related dependency. Here, we compare KRAS and EGFR status in NSCLC versus (vs) CRC in the large Response Genetics (RGI) database. Methods: KRAS MTs were assessed in 2,693 specimens (838 CRC and 1165 NSCLC) using DNA from microdissected formalin fixed paraffin embedded tumor specimens. EGFR MTs were determined in 649 NSCLCs. A PCR approach with direct sequencing was used to detect 7 different base substitutions in KRAS and 28 EGFR MTs. Statistics: Fisher's Exact Test. Results: KRAS MTs were found in 39% of CRC and 23% of NSCLC specimens (p < 0.001). EGFR MTs were identified in 16% of NSCLCs. In 447 specimens with both assays, EGFR and KRAS MTs were largely exclusive; 4 specimens harbored both (p < 0.001). The ratio of base transversions to transitions was 3.27 in NSCLC versus 0.77 in CRC (p < 0.001). With regard to histology, in NSCLC KRAS MTs were observed in 28.7% (186/649) of adenocarcinomas vs 3.5% (7/198) of squamous cell carcinomas (p < 0.001). Tobacco carcinogenesis-associated G>T transversions (codon 12 GGT>TGT plus GGT>GTT) comprised 61% of KRAS MTs in NSCLC vs 39% in CRC (p < 0.001). The GGT>GAT transition was most prevalent in CRC at 33.3%. Conclusions: This analysis of the RGI database demonstrates differences between NSCLC vs CRC in both incidence and type of KRAS MTs. Increased frequencies of DNA KRAS transversions were seen in NSCLC, likely linked to tobacco exposure. Distinct MT patterns and biologic function may contribute to differences in predictive value for cetuximab therapy between NSCLC and CRC. Author Disclosure Employment or Leadership Position Consultant or Advisory Role Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Response Genetics Amgen, AstraZeneca, Biodesix, Boehringer Ingelheim, Bristol-Myers Squibb, Genentech, Genoptix, GlaxoSmithKline, Lilly, Merck, Novartis, Pfizer, Response Genetics, sanofi-aventis Response Genetics DxS, Response Genetics Abbott Laboratories, Bristol-Myers Squibb, Genentech, Lilly, Merck, Novartis, OSI, Pfizer

Predictive value of EGFR and HER2 overexpression in advanced non-small-cell lung cancer

Epidermal growth factor receptor (EGFR) and HER2 are cell surface receptor tyrosine kinases (TKs) that transduce growth signals through dimerization with HER family receptors. The heterodimerization of EGFR with HER2 induces a more potent activation of EGFR TK than does EGFR homodimerization. When tumor cells overexpress both EGFR and HER2, they exhibit aggressive tumor cell growth, owing to the increased potential for EGFR/HER2 heterodimerization and signaling. Gefitinib and erlotinib are EGFR TK inhibitors (EGFR TKIs) and have antitumor activity in 8-18% of patients with advanced non-small-cell lung cancer (NSCLC). Certain patient subsets are particularly responsive to EGFR TKIs. Analyses of biomarkers from patients in clinical studies of EGFR TKIs show correlations between objective tumor response and EGFR overexpression, as detected by immunohistochemistry and increased gene copy number measured by fluorescence in situ hybridization analysis. Furthermore, NSCLC tumors that overexpress both EGFR and HER2 are more sensitive to EGFR TKIs than are tumors that overexpress EGFR but are HER2 negative. Therefore, the measurement of EGFR and HER2 protein expression and the gene copy number in NSCLC tumors may have a prognostic value in NSCLC and a predictive value for identifying patients likely to benefit from an EGFR TKI. These considerations suggest that the simultaneous inhibition of EGFR and HER2 may warrant further study in patients with NSCLC.

Primary Tumor Standardized Uptake Value (SUVmax) Measured on Fluorodeoxyglucose Positron Emission Tomography (FDG-PET) is of Prognostic Value for Survival in Non-small Cell Lung Cancer (NSCLC): A Systematic Review and Meta-Analysis (MA) by the European Lung Cancer Working Party for the IASLC Lung Cancer Staging Project

The 2-[18F]-fluoro-2-deoxy-d-glucose positron emission tomography is an imaging tool for assessing clinical tumor, node, metastasis in non-small cell lung cancer (NSCLC). Primary tumor standardized uptake value (SUV) has been studied as a potential prognostic factor for survival. However, the sample sizes are limited leading to conduct a meta-analysis to improve the precision in estimating its effect. We performed a systematic literature search. For each publication, we extracted an estimate of the hazard ratio (HR) for comparing patients with a low and a high SUV and we aggregated the individual HRs into a combined HR, using a random-effects model. We found 13 eligible studies dedicated to NSCLC. Most of them included patients with stages I to III/IV and used a SUV assessment corrected for body weight. Number of patients ranged from 38 to 315 (total: 1474); 11 studies identified a high SUV as a poor prognostic factor for survival although two studies found no significant correlation between SUV and survival. SUV measurement and SUV threshold for defining high SUV were study dependent, eight studies looked for a so-called best cutoff (maximizing the logrank test statistic) without adjusting the p value for multiplicity. Overall, the combined HR for the 13 reports was 2.27 (95% confidence interval [CI]: 1.70-3.02); excluding the studies proposing a "best" cutoff, it was 2.08 (95% CI: 1.431-3.04). Our meta-analysis suggests that the primary tumor SUV measurement has a prognostic value in NSCLC; these results should be confirmed in a meta-analysis on individual patients' data.

Prognostic Model of Stage II Non-Small Cell Lung Cancer by a Discriminant Analysis of the Immunohistochemical Protein Expression

We aimed to identify the key proteins that influence the prognosis of non-small cell lung cancer (NSCLC) using protein expression profiles of previously known prognostic markers. Thirty-one cases of Stage II NSCLC with 5-year follow-up data were selected. Tissue microarrays (TMA) and immunohistochemistry were used to make protein expression profiles of 18 previously reported immunohistochemical prognostic markers and their value in NSCLC was statistically re-evaluated by a discriminant analysis. For the discriminant analysis using marker protein expression profiles, we selected three significant markers, TTF-1, RCAS1 and c-MET, to evaluate each patient's 5-year survival. The requested discriminant function was V = -1.08754 x (RCAS1 score) - 0.83174 x (TTF1 score) + 0.55204 x (cMET score) + 5.46972, and V = 0 served as a cut-off point. The correctness for evaluating a patient's 5-year survival by a discriminant analysis was 87.1%. A discriminant analysis is thus considered to be a useful statistical method for analyzing the protein expression profiles obtained by combined TMA and immunohistochemical techniques using archival NSCLC tissues. However, the sample size and selection of the marker protein depending on the histology greatly influence the results of a NSCLC study.

Biological and prognostic role of acid cysteine proteinase inhibitor (ACPI, cystatin A) in non-small-cell lung cancer

Background: Acid cysteine protease inhibitor (ACPI) is an intracellular protein, often linked to neoplastic changes in epithelium and thought to have an inhibitory role in malignant transformation. Aim: To analyse...