Biomarker Commons

Next-generation sequencing identifies novel microRNAs in peripheral blood of lung cancer patients.

Mol Biosyst. 2011 Dec;7(12):3187-99

Authors: Keller A, Backes C, Leidinger P, Kefer N, Boisguerin V, Barbacioru C, Vogel B, Matzas M, Huwer H, Katus HA, Stähler C, Meder B, Meese E

Abstract
MicroRNAs (miRNAs) are increasingly envisaged as biomarkers for various tumor and non-tumor diseases. MiRNA biomarker identification is, as of now, mostly performed in a candidate approach, limiting discovery to annotated miRNAs and ignoring unknown ones with potential diagnostic value. Here, we applied high-throughput SOLiD transcriptome sequencing of miRNAs expressed in human peripheral blood of patients with lung cancer. We developed a bioinformatics pipeline to generate profiles of miRNA markers and to detect novel miRNAs with diagnostic information. Applying our approach, we detected 76 previously unknown miRNAs and 41 novel mature forms of known precursors. In addition, we identified 32 annotated and seven unknown miRNAs that were significantly altered in cancer patients. These results demonstrate that deep sequencing of small RNAs bears high potential to quantify miRNAs in peripheral blood and to identify previously unknown miRNAs serving as biomarker for lung cancer.

PMID: 22027949 [PubMed - indexed for MEDLINE]

Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and -independent approaches.

J Thorac Oncol. 2012 Feb;7(2):306-15

Authors: Krebs MG, Hou JM, Sloane R, Lancashire L, Priest L, Nonaka D, Ward TH, Backen A, Clack G, Hughes A, Ranson M, Blackhall FH, Dive C

Abstract
INTRODUCTION: Epithelial circulating tumor cells (CTCs) are detectable in patients with non-small cell lung cancer (NSCLC). However, epithelial to mesenchymal transition, a widely reported prerequisite for metastasis, may lead to an underestimation of CTC number. We compared directly an epithelial marker-dependent (CellSearch) and a marker-independent (isolation by size of epithelial tumor cells [ISET]) technology platform for the ability to identify CTCs. Molecular characteristics of CTCs were also explored.
METHODS: Paired peripheral blood samples were collected from 40 chemonäive, stages IIIA to IV NSCLC patients. CTCs were enumerated by Epithelial Cell Adhesion Molecule-based immunomagnetic capture (CellSearch, Veridex) and by filtration (ISET, RareCell Diagnostics). CTCs isolated by filtration were assessed by immunohistochemistry for epithelial marker expression (cytokeratins, Epithelial Cell Adhesion Molecule, epidermal growth factor receptor) and for proliferation status (Ki67).
RESULTS: CTCs were detected using ISET in 32 of 40 (80%) patients compared with 9 of 40 (23%) patients using CellSearch. A subpopulation of CTCs isolated by ISET did not express epithelial markers. Circulating tumor microemboli (CTM, clusters of ≥ 3 CTCs) were observed in 43% patients using ISET but were undetectable by CellSearch. Up to 62% of single CTCs were positive for the proliferation marker Ki67, whereas cells within CTM were nonproliferative.
CONCLUSIONS: Both technology platforms detected NSCLC CTCs. ISET detected higher numbers of CTCs including epithelial marker negative tumor cells. ISET also isolated CTM and permitted molecular characterization. Combined with our previous CellSearch data confirming CTC number as an independent prognostic biomarker for NSCLC, we propose that this complementary dual technology approach to CTC analysis allows more complete exploration of CTCs in patients with NSCLC.

PMID: 22173704 [PubMed - indexed for MEDLINE]

ΔNp63 (p40) and thyroid transcription factor-1 immunoreactivity on small biopsies or cellblocks for typing non-small cell lung cancer: a novel two-hit, sparing-material approach.

J Thorac Oncol. 2012 Feb;7(2):281-90

Authors: Pelosi G, Fabbri A, Bianchi F, Maisonneuve P, Rossi G, Barbareschi M, Graziano P, Cavazza A, Rekhtman N, Pastorino U, Scanagatta P, Papotti M

Abstract
INTRODUCTION: Diagnosing non-small cell lung cancer on biopsy/cellblock samples by morphology may be demanding. As sparing material for molecular testing is mandatory, a minimalist immunohistochemistry (IHC)-based diagnostic approach is warranted by means of novel, reliable, and easy-to-assess biomarkers.
METHODS: Forty-six consecutive biopsy/cellblock samples and the corresponding resection specimens (as the gold standard for morphology and IHC) from 30 adenocarcinomas (AD), 10 squamous carcinomas (SQC), 5 adenosquamous carcinomas (ADSQC), and 1 sarcomatoid carcinoma (SC) were IHC-evaluated for p40 [corresponding to nontransactivating ΔNp63 isoforms] and thyroid transcription factor-1 (TTF1) by semiquantitative assessment. For p40, also immunodecoration intensity was taken into account and dichotomized as strong or low.
RESULTS: Nonrandom and overlapping distributions of the relevant markers were found in biopsy/cellblock and surgical specimens, which closely correlated with each other and the diverse tumor categories, with no differences in area under curve-receiver-operating-characteristic curves for each marker between any two samples, including p40 and p63. Diagnostic combinations were p40-/TTF1+ or TTF1- for AD (where p40 was negative, apart from 5/30 AD showing at the best 1-2% tumor cells with low intensity); p40+/TTF1- (p40 strong and by far higher than 50%) for SQC; and p40+/TTF1+ or p40+/TTF1- (p40 strong and less than 50%) for ADSQC. The single SC case was p40-/TTF1-, suggesting glandular lineage. Practically, 41/46 (89%) tumors were correctly classified by IHC on small samples, including 30 AD, 10 SQC, 1/5 ADSQC, and no SC. Underdiagnosis of ADSQC was actually because of sampling error of biopsies/cellblocks rather than insufficient biomarker robustness, whereas underdiagnosis of SC was really because of the failure of either marker to highlight epithelial-mesenchymal transition.
CONCLUSIONS: This minimalist IHC-based model of p40 and TTF1 on biopsy/cellblock samples was effective to correctly subtype most cases of lung cancer.

PMID: 22071786 [PubMed - indexed for MEDLINE]

The cost-effectiveness of screening lung cancer patients for targeted drug sensitivity markers.

Br J Cancer. 2012 Mar 13;106(6):1100-6

Authors: Atherly AJ, Camidge DR

Abstract
BACKGROUND: New oncology drugs are being developed in conjunction with companion diagnostics with approval restricting their use to certain biomarker-positive subgroups. We examined the impact of different predictive biomarker screening techniques and population enrichment criteria on the cost-effectiveness of targeted drugs in lung cancer, using ALK and crizotinib to build the initial model.
METHODS: Health economic modeling of cost per Quality Adjusted Life Year was based on literature review and expert opinion. The modeled population represented advanced non-small cell lung cancer (NSCLC), eligible for predictive biomarker screening with prescribing restricted to biomarker-positive patients.
RESULTS: For assays costing $1400 per person, cost per quality-adjusted life year (QALY) gained for ALK screening all advanced NSCLC, excluding treatment cost, is $106,707. This falls to $4756 when only a highly enriched population is screened (increasing biomarker frequency from 1.6 to 35.9%). However, the same enrichment involves missing 56% patients who segregate within the unscreened group. Cheaper screening tests that miss some true positives can be more cost-effective if proportional reductions in cost exceed proportion of subjects missed. Generic modeling of idealised screening assays, including treatment cost, reveals a dominant effect of screening cost per person at low biomarker frequencies. Cost-effectiveness of <$100,000 per QALY gained is not achievable at biomarker frequencies <5% (with drug costs $1-5000 per month and screening costs $600-1400 per person).
INTERPRETATION: Cost-effectiveness of oncology drugs whose prescribing is restricted to biomarker-positive subgroups should address the cost of detecting marker-positive patients. The cost of screening dominates at low frequencies and strategies to improve cost-effectiveness based on the assay cost, drug cost and the group screened should be considered in these scenarios.

PMID: 22374459 [PubMed - indexed for MEDLINE]

Analysis of circulating angiogenic biomarkers from patients in two phase III trials in lung cancer of chemotherapy alone or chemotherapy and thalidomide.

Br J Cancer. 2012 Mar 13;106(6):1153-9

Authors: Young RJ, Tin AW, Brown NJ, Jitlal M, Lee SM, Woll PJ

Abstract
BACKGROUND: Thalidomide has potent anti-inflammatory and anti-angiogenic properties. It was evaluated in combination with chemotherapy in two randomised placebo-controlled trials in patients with small cell lung cancer (SCLC, n=724) and advanced non-small cell lung cancer (NSCLC, n=722). Neither study demonstrated an improvement in overall survival with the addition of thalidomide to chemotherapy. This study investigated circulating angiogenic biomarkers in a subset of these patients.
METHODS: Serial plasma samples were collected in a cohort of patients enrolled in these two trials (n=95). Vascular endothelial growth factor (VEGF), soluble truncated form of VEGF receptor-2 (sVEGFR-2), interleukin-8 (IL-8), tumour necrosis factor-α (TNF-α), basic fibroblast growth factor (bFGF) and soluble intercellular adhesion molecule-1 (sICAM-1) levels were measured by enzyme-linked immunosorbent assays. Results were correlated with patient clinical data including stage, response rate and progression-free survival (PFS).
RESULTS: Baseline biomarker levels were not significantly different between SCLC and NSCLC. For pooled treatment groups, limited stage SCLC was associated with lower baseline VEGF (P=0.046), sICAM-1 (P=0.008) and IL-8 (P=0.070) than extensive stage disease. Low baseline IL-8 was associated with a significantly improved PFS in both SCLC and NSCLC (P=0.028), and a greater reduction in IL-8 was associated with a significantly improved tumour response (P=0.035). Baseline angiogenic factor levels, however, did not predict response to thalidomide.
CONCLUSION: Circulating angiogenic biomarkers did not identify patients who benefited from thalidomide treatment.

PMID: 22353811 [PubMed - indexed for MEDLINE]

New testing for lung cancer screening.

Oncology (Williston Park). 2012 Feb;26(2):176-82

Authors: Tanner NT, Mehta H, Silvestri GA

Abstract
Lung cancer is the leading cause of cancer-related death worldwide. At the time of initial presentation, most patients are at an advanced stage of disease with an associated poor prognosis.Those diagnosed and treated at earlier stages have a significantly better outcome, with 5-year survival for stage I disease approaching 75%. Ideally, a screening strategy for lung cancer would detect disease at an earlier stage and allow for potential surgical cure in those with cancer and avoid unnecessary morbidity in those without. Until recently, there had not been a screening test that demonstrated a mortality reduction in lung cancer. The National Lung Cancer Screening Trial showed that screening high-risk persons with low-dose CT scanning could significantly reduce lung cancer mortality; however, there are difficulties in generalizing these results to the community. Several new testing techniques, including measurement of volatile organic compounds in exhaled breath, an airway epithelial gene expression biomarker, and serum sampling for antibodies to tumor-associated antigens, are currently being evaluated and may prove useful as part of a screening algorithm for lung cancer.

PMID: 22489353 [PubMed - indexed for MEDLINE]

Molecular biomarkers of lung carcinoma.

Front Biosci (Elite Ed). 2012;4:865-75

Authors: Vrabec Branica B, Gajovic S

Abstract
Lung carcinoma is still the leading cause of cancer deaths in men and women. There is a constantly increasing need to find molecular biomarkers for lung cancer which can be used for risk stratification, early detection, treatment selection, prognosis and monitoring for recurrence. Recent advances in imaging and improved bronchoscopic techniques have intensified interest in lung carcinoma screening techniques, especially in new molecular markers which can help cytopathologists to make a definitive diagnosis on very small specimens in non-invasive, non-expensive, simple and efficient manner. Several decades of intensive research have originated numerous potential lung carcinoma molecular biomarkers but only few turned out to be useful in clinic. The review describes types of biomarkers, sources and techniques for their identification in cancer diagnosis and therapy. A deep understanding of each biomarker will be a key to efficiently diagnose lung carcinomas and direct patients toward beneficial drugs based on individual patient profile.

PMID: 22201920 [PubMed - indexed for MEDLINE]

Expression of C4.4A in precursor lesions of pulmonary adenocarcinoma and squamous cell carcinoma.

Int J Cancer. 2012 Jun 1;130(11):2734-9

Authors: Jacobsen B, Santoni-Rugiu E, Illemann M, Kriegbaum MC, Laerum OD, Ploug M

Abstract
The protein C4.4A, a structural homologue of the urokinase-type plasminogen activator receptor, is a potential new biomarker in non-small cell lung cancer, with high levels of expression recently shown to correlate to poor survival of adenocarcinoma patients. In this study, C4.4A immunoreactivity in precursor lesions of lung squamous cell carcinoma and adenocarcinoma was investigated by stainings with a specific anti-C4.4A antibody. In the transformation from normal bronchial epithelium to squamous cell carcinoma, C4.4A was weakly expressed in basal cell hyperplasia but dramatically increased in squamous metaplasia. This was confined to the cell membrane and sustained in dysplasia, carcinoma in situ, and the invasive carcinoma. The induction of C4.4A already at the stage of hyperplasia could indicate that it is a marker of very early squamous differentiation, which aligns well with our earlier finding that C4.4A expression levels do not provide prognostic information on the survival of squamous cell carcinoma patients. In the progression from normal alveolar epithelium to peripheral adenocarcinoma, we observed an unexpected, distinct cytoplasmic staining for C4.4A in a fraction of atypical adenomatous hyperplasias, while most bronchioloalveolar carcinomas were negative. Likewise, only a fraction of the invasive adenocarcinomas was positive for C4.4A. With a view to the prognostic impact of C4.4A in adenocarcinoma patients, this finding might suggest that C4.4A could be an early biomarker for a possibly more malignant subtype of this disease.

PMID: 21792890 [PubMed - indexed for MEDLINE]

Genome structure-based screening identified epigenetically silenced microRNA associated with invasiveness in non-small-cell lung cancer.

Int J Cancer. 2012 Jun 1;130(11):2580-90

Authors: Watanabe K, Emoto N, Hamano E, Sunohara M, Kawakami M, Kage H, Kitano K, Nakajima J, Goto A, Fukayama M, Nagase T, Yatomi Y, Ohishi N, Takai D

Abstract
MicroRNA (miRNA) expression is frequently altered in human cancers. To search for epigenetically silenced miRNAs in non-small-cell lung cancer (NSCLC), we mapped human miRNAs on autosomal chromosomes and selected 55 miRNAs in silico. We treated six NSCLC cell lines with the DNA methylation inhibitor 5-aza-2'-deoxycytidine (5-aza-CdR) and determined the expressions of the 55 miRNAs. Fourteen miRNAs were decreased in the cancer cell lines and were induced after 5-aza-CdR treatment. After a detailed DNA methylation analysis, we found that mir-34b and mir-126 were silenced by DNA methylation. Mir-34b was silenced by the DNA methylation of its own promoter, whereas mir-126 was silenced by the DNA methylation of its host gene, EGFL7. A chromatin immunoprecipitation assay revealed H3K9me2 and H3K9me3 in mir-34b and EGFL7, and H3K27me3 in EGFL7. The overexpression of mir-34b and mir-126 decreased the expression of c-Met and Crk, respectively. The 5-aza-CdR treatment of lung cancer cell line resulted in increased mir-34b expression and decreased c-Met protein. We next analyzed the DNA methylation status of these miRNAs using 99 primary NSCLCs. Mir-34b and mir-126 were methylated in 41 and 7% of all the cases, respectively. The DNA methylation of mir-34b was not associated with c-Met expression determined by immunohistochemistry, but both mir-34b methylation (p = 0.007) and c-Met expression (p = 0.005) were significantly associated with lymphatic invasion in a multivariate analysis. The DNA methylation of mir-34b can be used as a biomarker for an invasive phenotype of lung cancer.

PMID: 21702040 [PubMed - indexed for MEDLINE]

Imaging of tumor viability in lung cancer: initial results using 23Na-MRI.

Rofo. 2012 Apr;184(4):340-4

Authors: Henzler T, Konstandin S, Schmid-Bindert G, Apfaltrer P, Haneder S, Wenz F, Schad L, Manegold C, Schoenberg SO, Fink C

Abstract
PURPOSE: 23Na-MRI has been proposed as a potential imaging biomarker for the assessment of tumor viability and the evaluation of therapy response but has not yet been evaluated in patients with lung cancer. We aimed to assess the feasibility of 23Na-MRI in patients with lung cancer.
MATERIALS AND METHODS: Three patients with stage IV adenocarcinoma of the lung were examined on a clinical 3 Tesla MRI system (Magnetom TimTrio, Siemens Healthcare, Erlangen, Germany). Feasibility of 23Na-MRI images was proven by comparison and fusion of 23Na-MRI with 1H-MR, CT and FDG-PET-CT images. 23Na signal intensities (SI) of tumor and cerebrospinal fluid (CSF) of the spinal canal were measured and the SI ratio in tumor and CSF was calculated. One chemonaive patient was examined before and after the initiation of combination therapy (Carboplatin, Gemcitabin, Cetuximab).
RESULTS: All 23Na-MRI examinations were successfully completed and were of diagnostic quality. Fusion of 23Na-MRI images with 1H-MRI, CT and FDG-PET-CT was feasible in all patients and showed differences in solid and necrotic tumor areas. The mean tumor SI and the tumor/CSF SI ratio were 13.3 ± 1.8 × 103 and 0.83 ± 0.14, respectively. In necrotic tumors, as suggested by central non-FDG-avid areas, the mean tumor SI and the tumor/CSF ratio were 19.4 × 103 and 1.10, respectively.
CONCLUSION: 23Na-MRI is feasible in patients with lung cancer and could provide valuable functional molecular information regarding tumor viability, and potentially treatment response.

PMID: 22351502 [PubMed - indexed for MEDLINE]

Interferon (alpha, beta and omega) receptor 2 is a prognostic biomarker for lung cancer.

Pathobiology. 2012;79(1):24-33

Authors: Tanaka S, Hattori N, Ishikawa N, Horimasu Y, Deguchi N, Takano A, Tomoda Y, Yoshioka K, Fujitaka K, Arihiro K, Okada M, Yokoyama A, Kohno N

Abstract
OBJECTIVES: It has been reported that the type I interferon receptor subunit, interferon (alpha, beta and omega) receptor 2 (IFNAR2), is overexpressed in several malignancies, primarily adenocarcinomas (ADCs); however, the biological significance of IFNAR2 in human lung cancer has not yet been studied.
METHODS: Immunohistochemical analysis of 113 surgically resected lung specimens was performed, and the results were evaluated in association with clinical variables, including survival. Serum concentrations of IFNAR2 were also determined by an enzyme-linked immunosorbent assay in 157 lung cancer patients and 164 healthy volunteers.
RESULTS: IFNAR2 overexpression was observed in all histological types of lung cancer examined. Furthermore, strong IFNAR2 expression was associated with shorter progression-free survival (PFS) and overall survival (OS) (p < 0.0001 and p = 0.0110, respectively) in non-small cell lung cancer patients. Multivariate analyses confirmed its independent prognostic value for PFS and OS (p < 0.0001 and p = 0.0222, respectively). IFNAR2 serum levels were also significantly higher in lung cancer patients than in healthy volunteers (p < 0.0001).
CONCLUSIONS: IFNAR2 overexpression was observed in various histological types of lung cancer, and appears to be associated with lung cancers that behave aggressively. The results of this study strongly support the potential of IFNAR2 to be a prognostic biomarker for lung cancer.

PMID: 22236545 [PubMed - indexed for MEDLINE]

XAF1 as a prognostic biomarker and therapeutic target in squamous cell lung cancer.

Chin Med J (Engl). 2011 Oct;124(20):3238-43

Authors: Chen YB, Shu J, Yang WT, Shi L, Guo XF, Wang FG, Qian YY

Abstract
BACKGROUND: X-linked inhibitor of apoptosis (XIAP)-associated factor 1 (XAF1) is a new tumor suppressor. Low expression of XAF1 is associated with poor prognosis of human cancers. However, the effect of XAF1 on lung cancer remains unknown. In this study, we investigated the expression of XAF1 and its role in squamous cell lung cancer.
METHODS: Cancer tissues, cancer adjacent tissues and normal lung tissues were collected from 51 cases of squamous cell lung cancer. The expression of XAF1 mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). The expression of XAF1 protein was determined by Western blotting and immunohistochemical staining. Ad5/F35-XAF1 virus was generated. Cell proliferation and apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and flow cytometry (FACS), respectively.
RESULTS: The levels of XAF1 protein and mRNA in cancer tissues were significantly lower than those in cancer adjacent and normal lung tissues (P < 0.05). The low expression of XAF1 was associated with tumor grade, disease stage, differentiation status and lymph node metastasis in squamous cell lung cancer patients. The restoration of XAF1 expression mediated by Ad5/F35-XAF1 virus significantly inhibited cell proliferation and induced apoptosis in a dose- and time-dependent manner.
CONCLUSION: XAF1 is a valuable prognostic marker in squamous cell lung cancer and may be a potential candidate gene for lung cancer therapy.

PMID: 22088514 [PubMed - indexed for MEDLINE]

RACK1 promotes non-small-cell lung cancer tumorigenicity through activating sonic hedgehog signaling pathway.

J Biol Chem. 2012 Mar 9;287(11):7845-58

Authors: Shi S, Deng YZ, Zhao JS, Ji XD, Shi J, Feng YX, Li G, Li JJ, Zhu D, Koeffler HP, Zhao Y, Xie D

Abstract
Non-small-cell lung cancer (NSCLC) is a deadly disease due to lack of effective diagnosis biomarker and therapeutic target. Much effort has been made in defining gene defects in NSCLC, but its full molecular pathogenesis remains unexplored. Here, we found RACK1 (receptor of activated kinase 1) was elevated in most NSCLC, and its expression level correlated with key pathological characteristics including tumor differentiation, stage, and metastasis. In addition, RACK1 activated sonic hedgehog signaling pathway by interacting with and activating Smoothened to mediate Gli1-dependent transcription in NSCLC cells. And silencing RACK1 dramatically inhibited in vivo tumor growth and metastasis by blocking the sonic hedgehog signaling pathway. These results suggest that RACK1 represents a new promising diagnosis biomarker and therapeutic target for NSCLC.

PMID: 22262830 [PubMed - indexed for MEDLINE]

Membrane proteomic analysis comparing squamous cell lung cancer tissue and tumour-adjacent normal tissue.

Cancer Lett. 2012 Jun 1;319(1):118-24

Authors: Li B, Chang J, Chu Y, Kang H, Yang J, Jiang J, Ma H

Abstract
Lung cancer is the leading cause of cancer-related deaths worldwide. Squamous cell carcinoma is one of the predominant histological subtypes of lung cancer. Detecting lung cancer at an early stage is essential for successful therapy and increasing survival. There are still no satisfactory biomarkers for the early detection of lung cancer. In this study, tumour tissue paired with tumour-adjacent normal bronchial epithelial tissue was obtained from patients with squamous cell lung carcinoma without metastasis. The proteins extracted from the cell membrane were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE) and were analysed with the Image Master two-dimensional platinum software. Twenty-five significantly different protein spots were selected and identified with matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). A total of 19 proteins were successfully identified. Twelve proteins were up-regulated, and seven proteins were down-regulated in the cancerous tissue compared with the tumour-adjacent normal tissue. One up-regulated protein and one down-regulated protein in squamous cell lung carcinoma were verified by Western blot analysis and RT-PCR; the results were consistent with the 2-DE analysis. In conclusion, membrane proteomics identified a number of candidate biomarker proteins that were differentially expressed between squamous cell lung cancer tissue and adjacent normal tissue. These biomarker candidates have the potential to elucidate the underlying pathogenesis of squamous cell lung cancer.

PMID: 22252117 [PubMed - indexed for MEDLINE]

[Relationship between the protein expression of ERCC1, BRCA, β-tubulin and K-ras and the efficacy and prognosis in advanced non-small cell lung cancer].

Zhonghua Zhong Liu Za Zhi. 2011 Mar;33(3):212-6

Authors: Zhang L, Liu T, Zhang JQ

Abstract
OBJECTIVE: To investigate the relationship between the expression of ERCC1, BRCA1, β-tubulin and K-ras and the clinical efficacy, prognosis in advanced non-small cell lung cancer treated by platinum-based chemotherapy.
METHODS: Expression of ERCC1, BRCA1, β-tubulin and K-ras proteins were detected by immunohistochemistry in 136 patients. The relation between gene protein expression and efficacy, prognosis was analyzed.
RESULTS: (1) The efficacy of chemotherapy (ORR) in the ERCC1 negative group was better than that in the ERCC1 positive group (38.6% vs.26.4%, P < 0.017). The median survival time in the ERCC1 negative group was longer than that in the ERCC1 positive group (15 months vs. 12 months, P < 0.05). There was no significant difference in PFS. (2) The clinical stage in the BRCA1 negative group was earlier than that of positive group. There was no significant difference in ORR in either the BRCA1 negative group or BRCA1 positive group. But both MST and PFS in the BRCA1 negative group was longer than that in the BRCA1 positive group (16 months vs. 9 months, P < 0.05 and 7 months vs. 7 months, P < 0.05), respectovely. (3) There were no significant differences between the β-tubulin negative group and positive group in MST, PFS and ORR. (4) The ORR in the K-ras negative group was better than that in the K-ras positive group. There was no significant difference between the K-ras negative and positive groups in MST and PFS. (5) ERCC1 protein was an independent prognostic factor determined by multivariate analysis.
CONCLUSIONS: Multi-biomarker detection may provide important predictive value for chemotherapy efficacy and prognosis in advanced NSCLC.

PMID: 21575522 [PubMed - indexed for MEDLINE]

Enzastaurin has anti-tumour effects in lung cancers with overexpressed JAK pathway molecules.

Br J Cancer. 2012 Feb 28;106(5):867-75

Authors: Shimokawa T, Seike M, Soeno C, Uesaka H, Miyanaga A, Mizutani H, Kitamura K, Minegishi Y, Noro R, Okano T, Yoshimura A, Gemma A

Abstract
BACKGROUND: Enzastaurin, an oral serine-threonine kinase inhibitor, was initially developed as an ATP-competitive selective inhibitor against protein kinase Cβ. However, the mechanism by which enzastaurin contributes to tumourigenesis remains unclear.
METHODS: We analysed the anti-tumour effects of enzastaurin in 22 lung cancer cell lines to ascertain the potential for enzastaurin-based treatment of lung cancer. To identify molecules or signalling pathways associated with this sensitivity, we conducted a gene, receptor tyrosine kinases phosphorylation and microRNA expression profiling study on the same set of cell lines.
RESULTS: We identified eight genes by pathway analysis of molecules having gene-drug sensitivity correlation, and used them to build a support vector machine algorithm model by which sensitive cell lines were distinguished from resistant cell lines. Pathway analysis revealed that the JAK/STAT signalling pathway was one of the main ones involved in sensitivity to enzastaurin. Overexpression of JAK1 was observed in the sensitive cells by western blotting. Simultaneous administration of enzastaurin and JAK inhibitor inhibited enzastaurin-induced cell growth-inhibitory effect. Furthermore, lentiviral-mediated JAK1-overexpressing cells were more sensitive to enzastaurin than control cells.
CONCLUSION: Our results suggested that the JAK1 pathway may be used as a single predictive biomarker for enzastaurin treatment. The anti-tumour effect of enzastaurin should be evaluated in lung cancer with overexpressed JAK pathway molecules.

PMID: 22333600 [PubMed - indexed for MEDLINE]

Plasma biomarkers distinguish non-small cell lung cancer from asthma and differ in men and women.

Cancer Genomics Proteomics. 2012 Jan;9(1):27-35

Authors: Izbicka E, Streeper RT, Michalek JE, Louden CL, Diaz A, Campos DR

Abstract
BACKGROUND: Lung cancer (LC) is the leading cause of deaths caused by cancer worldwide. A diagnostic test for LC is needed for monitoring high-risk populations.
PATIENTS AND METHODS: Fifty-seven markers were measured using multiplex immunoassays of plasma of patients with non-small cell lung cancer (NSCLC); (245 men, 114 women, 1 unknown), asthma (67 men, 111 women, 2 unknown) and of healthy controls (165 men, 122 women, 1 unknown). Mass spectrometry was used for biomarker discovery. A support vector machine (SVM) was used for data analysis.
RESULTS: When all biomarkers and both genders were co-analyzed, SVM classified NSCLC and asthma with an accuracy of 0.94. Restricting to NSCLC versus healthy using best subsets of variables (males: epidermal growth factor (EGF), interleukin-8 (IL-8), soluble Fas (sFas), matrix metalloproteinase-9 (MMP-9), plasminogen activator inhibitor-1 (PAI-1); females: EGF, soluble cluster of differentiation 40 (sCD40) ligand, IL-8) yielded sensitivity and specificity of 1. Expression of eleven mass spectrometric biomarkers differed between pathologies.
CONCLUSION: Significant inter-pathology and gender differences between biomarkers may improve diagnosis of LC.

PMID: 22210046 [PubMed - indexed for MEDLINE]

Association between 8-oxo-7,8-dihydroguanine excretion and risk of lung cancer in a prospective study.

Free Radic Biol Med. 2012 Jan 1;52(1):167-72

Authors: Loft S, Svoboda P, Kawai K, Kasai H, Sørensen M, Tjønneland A, Vogel U, Møller P, Overvad K, Raaschou-Nielsen O

Abstract
Oxidative damage to guanine (8-oxoGua) is one of the most abundant lesions induced by oxidative stress and documented mutagenic. 8-Oxoguanine DNA glycosylase 1 (OGG1) removes 8-oxoGua from DNA by excision. The urinary excretion of 8-oxoGua is a biomarker of exposure, reflecting the rate of damage in the steady state. The aim of this study was to investigate urinary 8-oxoGua as a risk factor for lung cancer. In a nested case-cohort design we examined associations between urinary excretion of 8-oxoGua and risk of lung cancer as well as potential interaction with the OGG1 Ser326Cys polymorphism in a population-based cohort of 25,717 men and 27,972 women aged 50-64 years with 3-7 years follow-up. We included 260 cases with lung cancer and a subcohort of 263 individuals matched on sex, age, and smoking duration for comparison. Urine collected at entry was analysed for 8-oxoGua by HPLC with electrochemical detection. There was no significant effect of smoking or OGG1 genotype on the excretion of 8-oxoGua. Overall the incidence rate ratio (IRR) (95% confidence interval) of lung cancer was 1.06 (0.97-1.15) per doubling of 8-oxoGua excretion. The association between lung cancer risk and 8-oxoGua excretion was significant among men [IRR: 1.17 (1.03-1.31)], never-smokers [IRR: 9.94 (1.04-94.7)], and former smokers [IRR: 1.19 (1.07-1.33)]. There was no significant interaction with the OGG1 genotype, although the IRR was 1.14 (0.98-1.34) among subjects homozygous for Cys326. The association between urinary 8-oxoGua excretion and lung cancer risk among former and never-smokers suggests that oxidative stress with damage to DNA is important in this group.

PMID: 22044660 [PubMed - indexed for MEDLINE]

Insulin-like growth factor II-messenger RNA-binding protein-3 and lung cancer.

Biotech Histochem. 2012 Jan;87(1):24-9

Authors: Findeis-Hosey JJ, Xu H

Abstract
Insulin-like growth factor II-messenger RNA-binding protein 3 (IMP3) is an oncofetal RNA-binding protein that promotes tumor cell proliferation by enhancing IGF-II protein synthesis and inducing cell adhesion and invasion by stabilizing CD44 mRNA. IMP3 expression has been studied in many human neoplasms with growing evidence that IMP3 is a biomarker of enhanced tumor aggressiveness. IMP3 expression has been correlated with a poorer phenotypic profile including increased risk of metastases and decreased survival. Only a few studies have examined IMP3 expression in lung cancers. We review here the literature concerning IMP3 expression in lung neoplasms, specifically adenocarcinoma, squamous cell carcinoma, and neuroendocrine tumors of the lung. IMP3 immunohistochemical expression was reported in 27-55% of cases of primary pulmonary adenocarcinoma and in 75-90% of cases of squamous cell carcinoma of the lung. In adenocarcinoma, IMP3 expression was reported to be correlated with more poorly differentiated histological grade, advanced stage of disease and lymph node metastases. IMP3 expression also may be a marker of high grade pre-invasive squamous lesions including high grade dysplasia and carcinoma in situ. In neuroendocrine tumors of the lung, IMP3 expression was expressed in all reported cases of large cell neuroendocrine carcinoma and small cell lung carcinoma, but expression was limited in carcinoid tumors. Overall, IMP3 appears to be a useful diagnostic marker for lung cancer pathology including for discriminating high grade neuroendocrine tumors and low grade carcinoids and for identifying high grade pre-invasive squamous lesions.

PMID: 21838610 [PubMed - indexed for MEDLINE]

Enhancer of zeste homolog 2 is a novel prognostic biomarker in nonsmall cell lung cancer.

Cancer. 2012 Mar 15;118(6):1599-606

Authors: Huqun , Ishikawa R, Zhang J, Miyazawa H, Goto Y, Shimizu Y, Hagiwara K, Koyama N

Abstract
BACKGROUND: Enhancer of zeste homolog 2 (EZH2) epigenetically silences many genes through the trimethylation of histone H3 lysine 27 and is implicated in tumor growth, invasion, and metastasis. However, its role in lung cancer has not been well characterized. The objective of the current study was to elucidate the role of EZH2 in nonsmall cell lung cancer (NSCLC) by investigating both clinical samples and cell lines.
METHODS: An immunohistochemical analysis of EZH2 expression was performed in samples from patients with stage I NSCLC to investigate the association of EZH2 expression levels with clinicopathologic variables. An in vitro cell growth assay and a Matrigel invasion assay also were conducted in the EZH2-expressing NSCLC cell lines A549 and H1299 after knocking down EZH2 expression by using an EZH2-specific short-hairpin RNA.
RESULTS: The immunohistochemical analysis classified stage I NSCLC samples (n = 106) into a negative EZH2 expression group (n = 40; 37.7%) and a positive EZH2 expression group (n = 66; 62.3%). Positive EZH2 expression was associated significantly with larger tumor size (P = .014). Kaplan-Meier survival analyses and log-rank tests demonstrated that patients whose samples were classified into the positive EZH2 expression group had a significantly shorter overall survival (P = .015). Experiments in the NSCLC cell lines revealed that the knockdown of EZH2 expression reduced the tumor growth rate and invasive activity.
CONCLUSIONS: The current results indicated that EZH2 promotes progression and invasion of NSCLC, and its expression is a novel prognostic biomarker in NSCLC.

PMID: 21837672 [PubMed - indexed for MEDLINE]