Prognosis Of Non-small Cell Lung Cancer Patients Research Articles

Expression and significance of Snail and Claudin-3 in non-small cell lung cancer

背景与目的上皮-间质转化（epithelial mesenchymal transition, EMT）是肿瘤浸润和转移的关键步骤，上皮细胞极性丧失是其主要标志，表现为Claudin等上皮标记丢失。锌指转录因子Snail是调控EMT的重要转录因子，近年来对肿瘤侵袭转移机制研究发现Snail能提高多种肿瘤的侵袭能力。本研究旨在利用组织芯片技术探讨转录因子Snail和紧密连接蛋白（Claudin-3）在非小细胞肺癌（non-small cell lung cancer, NSCLC）及其淋巴结转移灶中的表达和意义。方法分别采用免疫组织化学MaxVision法和EnVision法检测59例癌旁正常肺组织、302例NSCLC原发灶以及57例淋巴结转移灶中Snail和Claudin-3的表达。结果Snail在癌旁正常肺组织、NSCLC原发灶以及淋巴结转移灶中的表达逐渐增强，差异具有统计学意义（P < 0.05）；Claudin-3在癌旁正常肺组织、NSCLC原发灶以及淋巴结转移灶中的表达逐渐减弱，差异具有统计学意义（P < 0.05）。在NSCLC原发灶中，Snail和Claudin-3的表达与肿瘤组织学类型有关（P < 0.05）。Spearman等级相关分析显示Snail与Claudin-3的表达呈负相关（r=-0.178, P=0.002)。Kaplan-Meier生存分析显示肿瘤大小、组织学类型、病理分级、有无癌转移、TNM分期、Snail的表达以及Snail与Claudin-3的差异性表达影响NSCLC患者的术后生存时间（P < 0.05）。Cox回归分析提示肿瘤大小、组织学类型、病理分级、有无癌转移和TNM分期是影响NSCLC患者预后的独立危险因素（P < 0.05）。结论Snail和Claudin-3在NSCLC的浸润、转移中具有重要意义，有助于对NSCLC患者预后的评价。

High expression of heme oxygenase-1 is associated with tumor invasiveness and poor clinical outcome in non-small cell lung cancer patients

Heme oxygenase-1 (HO-1), a rate-limiting enzyme in heme catabolism, is known to play a role in the protection of cells against oxidative stress, inflammation, anomalous proliferation and apoptosis. As yet, the role of HO-1 expression in non-small cell lung cancer (NSCLC) development and metastasis remains unclear and insufficient data are available regarding its impact on the prognosis of NSCLC patients. Seventy NSCLC patients who underwent surgical resection were included in this HO-1 expression study and, concomitantly, clinical parameters were collected. Two lung adenocarcinoma cell lines (A549 and H441) were used to assess both invasive and migratory parameters in vitro. NSCLC patients with a high HO-1 expression ratio (tumor tissue/normal tissue) (> 1) exhibited a significantly poorer prognosis and a higher metastatic rate compared to those with a low HO-1 expression ratio (p < 0.05). The invasive and migratory abilities of A549 and H441 cells significantly increased after exogenous HO-1 over-expression and significantly decreased after siRNA-mediated HO-1 expression silencing. HO-1 up- and down-regulation also positively correlated with the expression of metastasis-associated proteins EGFR, CD147 and MMP-9. In addition, we found that HO-1 expression can be inhibited by PI3K and AKT inhibitors, but not by MAPK inhibitors. HO-1 is a poor prognostic NSCLC predictor and its over-expression may increase the metastatic potential of NSCLC. Based on our findings and those of others, HO-1 may be considered as a novel NSCLC therapeutic target.

Metallothionein 1F and 2A overexpression predicts poor outcome of non-small cell lung cancer patients

Metallothioneins (MT) are intracellular, low molecular weight proteins (6–7kDa) involved in binding of metal ions, scavenging of free radicals, cell proliferation and apoptosis and resistance to certain chemotherapeutics. Four basic families of MT proteins are distinguished: MT-I, MT-II, MT-III, MT-IV, within each of them different isoforms occur. The study aimed at examining the expression level of nine MT isoforms: MT-1A, -1B, -1E, -1F, -1G, -1H, -1X, MT-2A and MT-IV by using real-time PCR and MT-I/II expression by immunohistochemical (IHC) technique in 69 cases of non-small cell lung cancer (NSCLC) and 12 non-malignant lung tissues (NMLT) and to correlate them with patients clinicopathological data and Ki-67 antigen expression. Out of all the analyzed cases, 62 (89.9%) demonstrated an increased MT-I/II expression. MT-1B, 1F, -1G, -1H and MT-1X were significantly up-regulated, whereas MT-1E was significantly down-regulated in NSCLC as compared to NMLT. Only in two cases MT-IV mRNA expression was noted. Significant positive correlations were observed between each particular MT isoform expressions. Higher MT-1F and MT-1A mRNA expression was associated with larger primary tumor size (P=0.0362 and P<0.0001, respectively). Moreover, up-regulated MT-1F mRNA expression was associated with higher grade of malignancy of NSCLC (P=0.0085). Higher MT-1B mRNA expression was associated with squamocellular and adenocarcinoma subtype of NSCLC (P=0.0358). Univariate analysis showed, that up-regulated MT-1F and MT-2A mRNA predicted poor patients' survival (P=0.0206 and P=0.0097, respectively). The levels of MT-1F and MT-2A mRNA could be considered as new markers of poor prognosis of NSCLC patients.

Low Perioperative Serum Prealbumin Predicts Early Recurrence after Curative Pulmonary Resection for Non‐Small‐Cell Lung Cancer

Early recurrence after surgery for non-small-cell lung cancer (NSCLC) is often observed in spite of pathologically proven early-stage disease. The aim of this study was to identify biomarkers that might be useful in predicting postoperative early recurrence of lung cancer. In this study we evaluated the perioperative nutritional status of the patients by measuring the serum level of prealbumin and analyzed the correlation between this factor and early recurrence. Forty-four patients with NSCLC were enrolled in the study. Serum level of prealbumin was measured 5 days before and 7 days after surgery, respectively. For the patients who developed early recurrence, the perioperative serum prealbumin level was statistically significantly lower than those of the patients who did not develop recurrence (p<0.05). Furthermore, the patients with low prealbumin level showed statistically significantly poorer outcomes compared with the patients with higher prealbumin level (p<0.001). On the other hand, there was no correlation between the pathological stage and the serum prealbumin level. Multivariate analysis revealed that low perioperative serum prealbumin level could be an independent prognostic factor of poor outcome (hazard ratio, 10.1; 95% confidence interval, 2.8-35.5; p=0.0003). Low serum prealbumin level in the perioperative period is associated with a poorer prognosis in NSCLC patients and could serve as a marker for identifying patients at high risk, even at an early clinical stage.

1294P - Definitive Thoracic Chemoradiotherapy in Non-Small Cell Lung Cancer Patients with Solitary Brain Metastasis

ABSTRACT Background The aim of the study was to evaluate the impact of definitive thoracic chemoradiotherapy on outcomes in non-small cell lung cancer (NSCLC) patients with solitary brain metastasis. Materials and methods Fifty four medically fit NSCLC patients with isolated BM were retrospectively evaluated. Patients were staged with PET-CT besides conventional staging tools. TRT to a total dose of 66 Gy in 2 Gy fx was delivered with 2 cycles of concomitant cisplatin –based chemotherapy (CT) following surgery + 30 Gy whole-brain RT (WBRT) (n:18), 30 Gy WBRT + 15 Gy boost (n:22), or 30 Gy WBRT + stereotactic radiosurgery (SRS) (n:14) for their BM. Response evaluation was done according to EORTC RECIST criteria Results Pretreatment patients characteristics were as given in Table 1. Overall the treatment was well-tolerated and all patients received planned TRT and 2 cycles of CT. At a median follow up of 15.2 months (4.4-42.3), median overall, locoregional progression free and progression free survivals were 12.2, 8.5 and 5.9 months. Univariate analyses revealed that patients receiving WBRT + SRS for BM (P Conclusion Results of this study demonstrated that definitive T-CRT following WBRT + SRS for BM could improve poor prognosis in NSCLC patients with solitary BM. However, to make a firm conclusion, these findings should be validated by further prospective studies with larger cohorts. Pretreatment patients characteristics Characteristics Surgery + WBRT WBRT + BOOST WBRT + SRS P value Median age (range) 52.3 (33-68) 50.4 (32-66) 48.3 (38-62) 0.57 Sex [N,(%)] Female Male 11 (20.4) 7 (13.0) 14 (25.9) 8 (14.8) 10 (18.5) 4 (7.4) 0.82 Performance status [N,(%)] ECOG 0-1 ECOG 2 14 (25.9) 4 (7.4) 11 (20.4) 11 (20.4) 11 (20.4) 3 (5.6) 0.25 Histology [N,(%)] Squamous cell Adenocancer 11 (20.4) 7 (13.0) 14 (25.9) 8 (14.8) 5 (9.3) 9 (16.7) 0.22 T Stage T1 T2 T3 T4 3 (5.6) 4 (7.4) 5 (9.3) 6 (11.1) 3 (5.6) 4 (7.4) 3 (5.6) 12 (22.2) 3 (5.6) 2 (3.7) 7 (13.0) 2 (3.7) 0.20 N Stage N0 N1 N2 N3 6 (11.1) 5 (9.3) 6 (11.1) 1 (1.9) 2 (3.7) 7 (13.0) 5 (9.3) 8 (14.8) 2 (3.7) 2 (3.7) 7 (13.0) 3 (5.6) 0.10 ≥%5 weight loss [N,(%)] Yes No 9 (16.7) 9 (16.7) 12 (22.2) 10 (18.5) 5 (9.3) 9 (16.7) 0.54 Disclosure All authors have declared no conflicts of interest.

1663P - The Role of Hedgehog (HH) Signaling in the Prediction of Clinical Outcome for Advanced Non-Small Cell Lung Cancer (NSCLC)

ABSTRACT Background NSCLC lacks validated biomarkers to predict clinical outcome. The Hh pathway is critical for cell growth and differentiation. The aim of our study is to evaluate the role of the Hh signaling pathway in determining prognosis of NSCLC patients. Methods In this single-center prospective study, the expression of Hh-related molecules including Ptch1 and Gli1 (nuclear and cytoplasmic) was determined by immunohistochemistry in 35 histologic samples (biopsies and surgical specimens) of advanced NSCLC patients. All the neoplastic area included in the slides was considered and both cytoplasmic and nuclear stainings were evaluated, according to the positive neoplastic cells. The intensity of the staining was considered and scored as 0 (absent), 1+ (low), 2+ (medium) and 3+ (high). Results We analyzed 18 adenocarcinomas and 17 squamous cell carcinomas. Positivity of Gli1-cytoplasmic expression and Gli1-nuclear expression were expressed in adenocarcinoma at a significantly higher level and more frequently than compared to squamous cell carcinoma (p Conclusion At our best knowledge this is the first study investigating the role of HH in NSCLC patients. The results suggest that the Hh pathway might play a major prognostic role in NSCLC with significant differences between the histotype. Furthermore it might predict the efficacy of erlotinib as second-line treatment in patients with advanced lung adenocarcinoma. Disclosure All authors have declared no conflicts of interest.

Expression of TrkB and BDNF is associated with poor prognosis in non-small cell lung cancer

High expression levels of TrkB and BDNF are associated with aggressive malignant behavior in tumor cells and a poor prognosis in patients with various types of cancer. In this study, we aimed to identify the relationship between TrkB and BDNF expression and clinicopathological variables and prognosis in non-small cell lung cancer (NSCLC). We evaluated TrkB and BDNF expression in the tumor cells of 102 NSCLC patients by immunohistochemistry. Out of all clinicopathological factors examined, only vascular invasion was significantly correlated with TrkB (P=0.010) and BDNF (P=0.015) expression. TrkB-positive tumors had significantly worse disease-free survival (P=0.0094) and overall survival (P=0.0019) than TrkB-negative tumors, and TrkB expression was an independent prognostic factor for disease-free survival (HR 3.735, 95%C.I. 1.560–11.068, P=0.002) and overall survival (HR 4.335, 95%C.I. 1.534–15.963, P=0.004) in multivariate analysis. Finally, our analysis revealed that co-expression of TrkB and BDNF conferred poorer prognosis compared with overexpression of either protein alone. Our results indicate that expression of TrkB and BDNF is associated with poor prognosis in NSCLC patients.

Expression of Sox2 and Oct4 and Their Clinical Significance in Human Non-Small-Cell Lung Cancer

Sox2 and Oct4 are transcription factors with the characteristics of regulating self-renewal and differentiation of embryonic stem cell. The aim of this study was to detect the expression of Sox2 and Oct4 and analyze their clinical significance in human non-small-cell lung cancer (NSCLC). Expression of Sox2 and Oct4 were assayed in cancer tissues and their corresponding paracancerous tissues from 44 patients with NSCLC and 21 patients with benign tumors using immunohistochemistry, Western blot, reverse transcription polymerase chain reaction (RT-PCR). The correlation between the expression of Sox2 and Oct4 and tumor type, grade and prognosis and the utility of the two genes in discriminating between benign and malignant tumors were analyzed as well. The results showed that Sox2 and Oct4 positive staining was only seen in the nuclei of cancer cells but not in either the precancerous tissues or benign tumor tissues by immunohistochemistry (p < 0.01). Furthermore, in the lung cancer tissue, the positive rate for Sox2 and Oct4 was 70.5% and 54.5%, respectively. Meanwhile, clinicopathological correlations showed that the Oct4 expression level was significantly associated with poorer differentiation and higher TNM stage of the cancer (p < 0.05). Western blot and RT-PCR analysis showed similar results to immunohistochemistry. Follow-up analysis revealed that expression of Oct4 was significantly associated with poor prognosis of lung cancer. The conclusion is that Sox2 and Oct4 may act as the promising unit markers in directing NSCLC diagnosis and therapy. Also, Oct4 can be regarded as a novel predictor of poor prognosis for NSCLC patients undergoing resection.

Abstract 3395: TGFβ response signature in non-small cell lung carcinoma

Abstract Background: Transforming growth factor beta (TGFβ) is a cytokine involved in numerous cellular processes that include proliferation, migration and apoptosis. TGFβ is intriguing in that it plays a dual role in cancer; in some cases it functions as a tumor suppressor while in others it acts as a tumor promoter. Genetic alterations in the TGFβ signaling pathway have been reported in many cancers including breast, colon, gastric, lung, head and neck and pancreatic cancers. Correlations between a TGFβ induced gene expression and clinical outcomes have been described in breast and liver cancers, but not in non-small cell lung cancer (NSCLC). Because TGFβ has been shown to induce an epithelial-to-mesenchymal transition (EMT) in NSCLC that may lead to increased potential to invade and disseminate, we hypothesized that a TGFβ-induced gene expression signature might correlate with this transition and might predict prognosis in NSCLC patients. We therefore decided to attempt to identify gene expression changes induced by TGFβ that correlated with induced proliferation, migration, and EMT. We further investigated whether these TGFβ induced genes might be useful in the classification of NSCLC tumors. Methods: Ten different NSCLC cell lines were assessed for the ability of TGFβ-1 to induce migration, proliferation, and EMT. Cell lines were treated with TGFβ-1 for various time periods and the RNA collected for microarray analysis. Results: In nine out of the ten cell lines the TGFβ pathway was activated after treatment with TGFβ-1, but these lines differ in their TGFβ proliferation, migration and EMT responses. The H1944, H358, and A549 cell lines respond by shifting from an epithelial to a mesenchymal phenotype. Using the before and after treatment samples of these cell lines we looked for genes correlated with this transition. Initially, 632 probesets (534 genes) were identified that changed similarly in these 3 cell lines. Many of these changes seen after 120 hours of treatment were also seen at 48 hours but to a lesser degree. After Principal Component Analysis of the 534 genes in tumor sample a core set of 70 genes were retained and used to analyze lung cancer tumor samples. Using two independently derived sets of lung tumors the EMT signature could be used to identify 3 classes of tumors: tumors with high EMT gene expression, tumors with low EMT expression, and a distinct group of tumors that do not coordinately express the EMT genes identified in this study. Conclusion: Analysis of the differential responses of NSCLC cell lines to TGFβ-1 allows us to group NSCLC cell lines into at least three groups based on their response to TGFβ treatment. Using an EMT signature developed from the cell lines it appears that tumor samples can also be classified into three or more distinct groups that may differ in their in vivo responses to TGFβ-1. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3395. doi:1538-7445.AM2012-3395

Abstract 1734: Prognostic significance of combinations of RNA-dependent protein kinase and EphA2 biomarkers for NSCLC

Abstract RNA-dependent protein kinase (PKR) is an independent prognostic variable in patients with non-small cell lung cancer (NSCLC). In the present study, we investigated the correlation between PKR and 25 other biomarkers for NSCLC, identified the markers that could further improve the prognostic significance of PKR, and elucidated the mechanisms of interaction between these markers and PKR. Tissue microarray samples obtained from 218 lung cancer patients were stained with an anti-PKR antibody and antibodies against 25 biomarkers. Immunohistochemical expression was scored and used for Kaplan-Meier survival analysis. The interaction between PKR and EphA2 in NSCLC cell lines was examined. We found that PKR was associated with EphA2 and that the prognostic information regarding NSCLC provided by the combination of PKR and EphA2 (P/E) was significantly more accurate than that provided by either marker alone. The 5-year overall survival rate in PKRlow/EphA2high patients (20%) was significantly lower than that of PKRhigh/EphA2low patients (74%), PKRhigh/EphA2high patients (55%), and PKRlow/EphA2low patients (55%) (P &amp;lt; 0.0001). We also found that the PKR:EphA2 (P/E) ratio was significantly associated with prognosis (P &amp;lt; 0.0001). Univariate and multivariate Cox analyses revealed that this P/E combination or ratio was an independent predictor of overall survival. In addition, induction of PKR expression reduced EphA2 protein expression levels in NSCLC cell lines. PKR/EphA2 is a significant predictor of prognosis for NSCLC. PKR/EphA2 may be a promising approach to improving screening efficiency and predicting prognosis in NSCLC patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1734. doi:1538-7445.AM2012-1734

Abstract 5069: Genomic profiles of primary non-small cell lung cancer (NSCLC) xenograft tumors identify distinct gene signatures associated with histological subtypes

Abstract Xenografts established directly from patient tumors mirror closely the histology of the primary tumors. Therefore, primary tumor xenografts (PTXG) may serve as important preclinical models to evaluate novel anti-cancer drugs. We previously reported that the ability of resected tumors to engraft in NOD-scid mice is a strong predictor of relapse after surgery and poorer prognosis in NSCLC patients, and thus may represent biologically more aggressive cancers (Clin Cancer Res 2011;17:134-41). Genomic characterization of PTXG would help identify genetic aberrations that drive malignant oncogenic pathways in NSCLC. We characterized the somatic copy number alterations (CNA) of 36 PTGX, consisting of 15 adenocarcinoma (ADC), 18 squamous cell carcinoma (SCC), 2 large cell neuroendocrine carcinoma (LCNEC) and 1 large cell carcinoma (LC), along with 34 patient normal samples as controls using Illumina Omni-1 Quad SNP arrays. The gene expression profiles of the 36 PTGX were analyzed using Illumina Omni-1 Quad HT-12 v4 arrays. Histology-specific recurrent regions of CNA observed in PTGX are concordant with the published and publicly available primary NSCLC CNAs. We identified 1053 genes with somatic copy number gains and 932 genes with somatic copy number losses that distinguish between SCC and ADC. From integrative analysis of mRNA expression and somatic CNAs, we identified 325 genes specific to ADC and 2232 specific to SCC that are well correlated. Gene candidates that are deregulated in ADC include WRN, STK35, SIX1; and genes that are over-expressed in SCC include SOX2, RNF13, WNK1, PIK3CA, TFRC, TP63, PAK2 suggesting there is differential deregulation of signaling pathways between these two subtypes of lung cancer. We have identified candidate gene signatures that distinguish between ADC and SCC from PTXG, suggesting these xenograft models can provide a valuable resource to study cancer biology and preclinical drug target evaluation in vivo. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5069. doi:1538-7445.AM2012-5069

FoxM1 mediated resistance to gefitinib in non-smallcell lung cancer cells

Gefitinib is effective in only approximately 20% of patients with non-small-cell lung cancer (NSCLC), and the underlying mechanism remains unclear. FoxM1 is upregulated in NSCLC and associated with a poor prognosis in NSCLC patients. In this study, we examined the possible role of FoxM1 in gefitinib resistance and the related mechanisms. Gefitinib resistant human lung adenocarcinoma cell line SPC-A-1 and gefitinib-sensitive human lung mucoepidermoid carcinoma cell line NCI-H292 were used. mRNA and protein expression of FoxM1 and other factors were tested with quantitative RT PCR and Western blot analysis. RNA interference was performed to suppress FoxM1 expression in SPC-A-1 cells, and lentiviral infection was used to overexpress FoxM1 in H292 cells. MTT assay and flow cytometry were used to examine the proliferation and apoptosis of the cells. Treatment of SPC-A-1 cells with gefitinib (1 and 10 μmol/L) upregulated the expression of FoxM1 in time- and concentration-dependent manners, while gefitinib (1 μmol/L) downregulated in H292 cells. In SPC-A-1 cells treated with gefitinib (1 μmol/L), the expression of several downstream targets of FoxM1, including survivin, cyclin B1, SKP2, PLK1, Aurora B kinase and CDC25B, were significantly upregulated. Overexpression of FoxM1 increased the resistance in H292 cells, while attenuated FoxM1 expression restored the sensitivity to gefitinib in SPC-A-1 cells by inhibiting proliferation and inducing apoptosis. The results suggest that FoxM1 plays an important role in the resistance of NSCLC cells to gefitinib in vitro. FoxM1 could be used as a therapeutic target to overcome the resistance to gefitinib.

Prognostic significance of MCM7 expression in the bronchial brushings of patients with non-small cell lung cancer (NSCLC)

PurposeTo identify potential biomarkers for the prognosis of non-small cell lung cancer (NSCLC) patients by using bronchial brushing specimens. MethodsThe expression of MCM7, Ki67 and EGFR was evaluated in 494 NSCLC tissues and 174 bronchial brushings using immunohistochemical and immunocytochemical techniques. Associations between protein expression and clinico-pathologic parameters were assessed, and the impact on overall survival (OS) was analyzed. ResultsHigh expression of MCM7, Ki67 and EGFR was detected in 33.3%, 23.5% and 12.7% of tissues and in 52.4%, 52.7% and 20.6% of bronchial brushings, respectively. Expression of MCM7 and Ki67 was associated with squamous cell carcinoma (SCC) in both tissues and bronchial brushings (MCM7: P=0.0007, 0.00003; Ki67: P<0.00001, 0.00001). Overexpression of MCM7 in tumor tissues was detected more frequently in poorly differentiated tumors (P=0.0120) and non-bronchioloalveolar carcinomas (non-BACs) (P=0.0238). EGFR overexpression was observed in tissues of larger tumors (P=0.00004) and in bronchial brushings at later stage (P=0.0262). Kaplan–Meier curves indicated that patients with overexpression of MCM7 or Ki67 had a poorer OS compared to those with low expression for all stages (P<0.00001, 0.0233) and early-stages (P<0.00001, 0.0032). In particular, the patients with MCM7 overexpression in bronchial brushings had a poorer prognosis (P=0.0045). Multivariate Cox regression analysis showed that MCM7 was an independent prognostic indicator both in tissue samples and bronchial brushings. ConclusionsOur data suggest that MCM7 and Ki67 in tumor tissues may be potential markers of a poor prognosis for NSCLC patients. MCM7 in bronchial brushings also showed an independent prognostic value, which may be useful when biopsies are unavailable.

Association of Cytidine Deaminase and Xeroderma Pigmentosum Group D Polymorphisms with Response, Toxicity, and Survival in Cisplatin/Gemcitabine-Treated Advanced Non-small Cell Lung Cancer Patients

Selecting patients according to key genetic characteristics may help to tailor chemotherapy and optimize the treatment in non-small cell lung cancer (NSCLC). Genetic variations in drug metabolism may affect the clinical response, toxicity, and prognosis of NSCLC patients treated with cisplatin/gemcitabine-based therapy. We evaluated seven single-nucleotide polymorphisms of six genes CDA Lys27Gln (A/C); CDA C435T; ERCC1 C118T; XRCC3 Thr241Met (C/T); XPD Lys751Gln (A/C); P53 Arg72Pro (G/C), and RRM1 C524T in 192 chemotherapy-naive patients with advanced NSCLC treated with cisplatin/gemcitabine-based regimen by TaqMan probe-based assays with 7300 Real-Time PCR System, using genomic DNA extracted from blood samples. The CDA 435 T/T genotype was significantly associated with better response (p = 0.03). The CDA 435 C/T genotype was associated with a significantly increased risk of nonhematological toxicity of grade ≥3 (p = 0.02) after adjusting for performance status, age, and type of treatment regimen. In the multivariate Cox model, the XPD 751 C/C genotype was a significant prognostic factor of longer progression-free survival (p = 0.006). Our data suggest polymorphic variations of drug metabolic gene were associated with response and toxicity of cisplatin/gemcitabine-based therapy and progression-free survival of patients with advanced NSCLC.

Combined Effect of Genetic Polymorphisms in P53, P73, and MDM2 on Non-small Cell Lung Cancer Survival

Multiple biologically relevant polymorphisms may have more accurate prediction of cancer prognosis compared with single polymorphism because of the modest effect. This study investigated whether the functional polymorphisms in P53 pathway genes, P53 Arg72Pro (rs1042522), P73 G4C14-to-A4T14 (rs2273953 and rs1801173), and MDM2 T309G (rs2279744), alone or in combination, affect survival in advanced non-small cell lung cancer (NSCLC) patients. A total of 199 stage III-IV NSCLC patients with platinum-based chemotherapy were recruited between 2002 and 2004. Associations between genotypes and survival were assessed using Kaplan-Meier method. Cox proportional hazard models were performed to identify significant variables. During the median 26.5 months of follow-up, the P53 Pro/Pro genotype was strongly associated with shorter overall survival compared with the Arg/Arg genotype (12.0 versus 20.0 months; log-rank p = 0.002; hazard ratio = 1.86; 95% confidence interval [CI], 1.15-3.02). Pairwise combination analysis showed that patients carrying the variant P53 Pro/Pro-P73 GC/GC or P53 Pro/Pro-MDM2 GG genotypes had survival time only half of that for those carrying the wild-type genotypes, with hazard ratio being 2.47 (95% CI, 1.20-5.10) and 2.00 (95% CI, 1.15-3.46), respectively. Furthermore, a combined effect was seen with survival time being gradually shorter with increasing number of unfavorable genotypes in these three genes (p(trend) = 0.039), indicating a gene-dose effect in association with survival. These findings suggest that genetic polymorphisms in the P53 pathway may be promising biomarkers for individualized chemotherapy and prognosis of NSCLC patients.

TNFRSF1B +676 T&gt;G polymorphism predicts survival of non-Small cell lung cancer patients treated with chemoradiotherapy

BackgroundThe dysregulation of gene expression in the TNF-TNFR superfamily has been involved in various human cancers including non-small cell lung cancer (NSCLC). Furthermore, functional polymorphisms in TNF-α and TNFRSF1B genes that alter gene expression are likely to be associated with risk and clinical outcomes of cancers. However, few reported studies have investigated the association between potentially functional SNPs in both TNF-α and TNFRSF1B and prognosis of NSCLC patients treated with chemoradiotherapy.MethodsWe genotyped five potentially functional polymorphisms of TNF-α and TNFRSF1B genes [TNF-α -308 G>A (rs1800629) and -1031 T>C (rs1799964); TNFRSF1B +676 T>G (rs1061622), -1709A>T(rs652625) and +1663A>G (rs1061624)] in 225 NSCLC patients treated with chemoradiotherapy or radiotherapy alone. Kaplan-Meier survival analysis, log-rank tests and Cox proportional hazard models were used to evaluate associations between these variants and NSCLC overall survival (OS).ResultsWe found that the TNFRSF1B +676 GG genotype was associated with a significantly better OS of NSCLC (GG vs. TT: adjusted HR = 0.38, 95% CI = 0.15-0.94; GG vs. GT/TT: adjusted HR = 0.35, 95% CI = 0.14-0.88). Further stepwise multivariate Cox regression analysis showed that the TNFRSF1B +676 GG was an independent prognosis predictor in this NSCLC cohort (GG vs. GT/TT: HR = 0.35, 95% CI = 0.14-0.85), in the presence of node status (N2-3 vs. N0-1: HR = 1.60, 95% CI = 1.09-2.35) and tumor stage (T3-4 vs. T0-2: HR = 1.48, 95% CI = 1.08-2.03).ConclusionsAlthough the exact biological function for this SNP remains to be explored, our findings suggest a possible role of TNFRSF1B +676 T>G (rs1061622) in the prognosis of NSCLC. Further large and functional studies are needed to confirm our findings.

Prognostic significance of RIN1 gene expression in human non-small cell lung cancer

Ras interaction/interference 1 (RIN1), originally identified as a Ras effector protein, has been implicated in tumorigenesis and development of human cancers. The aim of this study was to detect RIN1 expression in human non-small cell lung cancer (NSCLC) and to analyze its association with prognosis of NSCLC patients. Quantitative real-time RT-PCR was performed to examine the expression of RIN1 mRNA in 25 cases of NSCLC and corresponding non-tumor tissue samples. Immunohistochemistry was performed to detect the expression of RIN1 in 90 NSCLC tissues. We found that the expression levels of RIN1 mRNA in NSCLC tissues were significantly higher than those in corresponding non-tumor tissues. High-level RIN1 expression was observed in 53.3% (48 of 90 cases), and correlated with poor tumor differentiation (P=0.024), TNM stage (P=0.032), and lymph node metastasis (P=0.018). Patients with high expression levels of RIN1 showed lower overall survival rate than those with low expression levels (P=0.033). Multivariate analysis showed that high RIN1 protein expression was an independent prognostic factor for NSCLC patients (P=0.021). Our study suggests that over-expression of RIN1 may play an important role in the progression of NSCLC and RIN1 expression may offer a valuable marker for predicting the outcome of patients with NSCLC.

Correlation between expression of HIF-2α and OCT-4 and prognosis of NSCLC

To investigate the expression and significance of hypoxia inducible factor-2α (HIF-2α) and transcription factor OCT-4 in non-small cell lung cancer (NSCLC), and evaluate their roles in the prognosis of NSCLC. Tissues from 51 cases of NSCLC were collected and immunohistochemistry (SP method) was used to detect the expression of HIF-2α and OCT-4 proteins. The correlation between the protein expression and the prognosis of NSCLC was analyzed. The positive rates of HIF-2α and OCT-4 expression in the NSCLC were 52.9% and 72.5%, respectively. There was significant relation between the expression of HIF-2α and OCT-4 (r=0.514,P<0.01). High expression of them revealed poor prognosis for NSCLC patients characterized with a bad overall survival(P<0.05). There is a negative corelation between the expression of HIF-2α and OCT-4 and the prognosis of NSCLC. Combined examination of HIF-2α and OCT-4 expression might be an important biomarker for NSCLC prognosis.

The expression and function of miRNA-451 in non-small cell lung cancer

miRNAs are small, non-coding RNAs that negatively regulate gene expression at the post-transcriptional and translational levels. miRNA-451 was previously reported to be down-regulated in gastric and colorectal cancers. Here, we showed that miRNA-451 expression decreased in non-small cell lung cancer (NSCLC) tissues and that its expression was negatively associated with lymph node metastasis, the stage of TNM classification and poor prognosis of NSCLC patients. Moreover, significantly different miRNA-451 expression levels were found between smoking and non-smoking patients. The overexpression of miRNA-451 inhibited cell cycle progression, cellular migration and the invasive ability of NSCLC cells. Increased miRNA-451 expression also promoted anoikis of NSCLC cells. Together, these data suggested that aberrantly expressed miRNA-451 may be associated with the development of NSCLC.

Prognostic significance of serum miRNA‐21 expression in human non‐small cell lung cancer

Deregulation of microRNAs (miRNAs) plays important roles in tumor progression. The aim of this study was to investigate miR-21 expression in serum of non-small cell lung cancer (NSCLC) and its correlation with prognosis of NSCLC patients. Dysregulated miRNAs in NSCLC serum were identified by microarray. MiR-21 expression in NSCLC and control serum was detected by TaqMan RT-PCR assay. The correlation of serum miR-21 with clinicopathological factors of NSCLC patients was analyzed. Furthermore, the prognostic significance of serum miR-21 was analyzed by using Kaplan-Meier curves with log-rank tests and a Cox proportional hazard model. The level of miR-21 expression was higher in NSCLC serum samples than in control serum samples (P < 0.01). High serum miR-21 was significantly correlated with tumor-node metastases stage and lymph node metastasis of NSCLC patients (P = 0.016 and 0.026, respectively). The 3-year actuarial overall survival rates in NSCLC patients with high serum miR-21 expression (39.8%) was significantly shorter than those with low serum miR-21 expression (58.2%; P < 0.001). Furthermore, univariate and multivariate analyses for overall survival showed that serum miR-21 expression was an independent prognostic factor for NSCLC patients (P = 0.015, RR = 2.01, 95% CI: 1.78-3.26). Serum miR-21 expression might be useful as a prognostic marker for NSCLC patients.