Genetic Modifiers of Progression-Free Survival in Never-Smoking Lung Adenocarcinoma Patients Treated with First-Line Tyrosine Kinase Inhibitors.

RATIONALE: Patients with non-small cell lung cancer (NSCLC) with mutated epidermal growth factor receptor (EGFR) are relatively sensitive to EGFR-tyrosine kinase inhibitor (TKI) treatment and have longer progression-free survival (PFS) when treated with EGFR-TKI compared with platinum-based chemotherapy. However, many patients with advanced NSCLC who have mutated EGFR do not respond to first-line EGFR-TKI treatment and still have shorter PFS. OBJECTIVES: The aim of this study was to identify genetic variants associated with PFS among patients with lung adenocarcinoma who were treated with first-line EGFR-TKIs. METHODS: A genome-wide association study on PFS was performed in never-smoking women diagnosed with lung adenocarcinoma and who were treated with first-line EGFR-TKIs (n = 128). Significant single-nucleotide polymorphisms (SNPs) were selected for follow-up association analysis (n = 198) and for replication assay in another independent cohort (n = 153). MEASUREMENTS AND MAIN RESULTS: We identified SNPs at 4q12 associated with PFS at genome-wide significance (P < 10-8) and with an estimated hazard ratio of more than 4. This association was also replicated in a larger but similar cohort and in an independent NSCLC cohort. Follow-up functional analyses showed that these SNPs were associated with the expression of EGFR, which encodes the TKI target, and with a nearby gene neuromedin-U, which encodes a G protein-coupled receptor ligand known to be involved in the progression of NSCLC. Considering these as possible prognostic biomarkers for the treatment of patients with late-stage lung cancer, we found that these SNPs were not associated with EGFR mutation status or with polymorphism of the Bcl2-interacting mediator of cell death gene. CONCLUSIONS: Genetic variants in 4q12 merit further investigation to assess their potential as pharmacogenomic predictors for and to understand the biology underlying its influence on PFS in patients treated with TKI therapy.

The database of chromosome imbalance regions and genes resided in lung cancer from Asian and Caucasian identified by array-comparative genomic hybridization.

BACKGROUND: Cancer-related genes show racial differences. Therefore, identification and characterization of DNA copy number alteration regions in different racial groups helps to dissect the mechanism of tumorigenesis. METHODS: Array-comparative genomic hybridization (array-CGH) was analyzed for DNA copy number profile in 40 Asian and 20 Caucasian lung cancer patients. Three methods including MetaCore analysis for disease and pathway correlations, concordance analysis between array-CGH database and the expression array database, and literature search for copy number variation genes were performed to select novel lung cancer candidate genes. Four candidate oncogenes were validated for DNA copy number and mRNA and protein expression by quantitative polymerase chain reaction (qPCR), chromogenic in situ hybridization (CISH), reverse transcriptase-qPCR (RT-qPCR), and immunohistochemistry (IHC) in more patients. RESULTS: We identified 20 chromosomal imbalance regions harboring 459 genes for Caucasian and 17 regions containing 476 genes for Asian lung cancer patients. Seven common chromosomal imbalance regions harboring 117 genes, included gain on 3p13-14, 6p22.1, 9q21.13, 13q14.1, and 17p13.3; and loss on 3p22.2-22.3 and 13q13.3 were found both in Asian and Caucasian patients. Gene validation for four genes including ARHGAP19 (10q24.1) functioning in Rho activity control, FRAT2 (10q24.1) involved in Wnt signaling, PAFAH1B1 (17p13.3) functioning in motility control, and ZNF322A (6p22.1) involved in MAPK signaling was performed using qPCR and RT-qPCR. Mean gene dosage and mRNA expression level of the four candidate genes in tumor tissues were significantly higher than the corresponding normal tissues (P<0.001~P=0.06). In addition, CISH analysis of patients indicated that copy number amplification indeed occurred for ARHGAP19 and ZNF322A genes in lung cancer patients. IHC analysis of paraffin blocks from Asian Caucasian patients demonstrated that the frequency of PAFAH1B1 protein overexpression was 68% in Asian and 70% in Caucasian. CONCLUSIONS: Our study provides an invaluable database revealing common and differential imbalance regions at specific chromosomes among Asian and Caucasian lung cancer patients. Four validation methods confirmed our database, which would help in further studies on the mechanism of lung tumorigenesis.

An E3 ubiquitin ligase: c-Cbl: a new therapeutic target of lung cancer.

BACKGROUND: Casitas B-lineage lymphoma (Cbl) is an E3 ubiquitin ligase of many tyrosine kinase receptors. The authors previously detected c-Cbl mutation and low protein expression in non-small cell lung cancer (NSCLC). Therefore, it was hypothesized that overexpression of wild-type c-Cbl (c-Cbl WT) exhibits tumor growth inhibition. METHODS: Wound healing and transwell assays were conducted to examine cell motility after c-Cbl WT transfection in NSCLC cell lines. The cell cycle was investigated by flow cytometry. A549 and H1299-Luc c-Cbl WT-transfected xenografts and experimental metastasis models were performed to investigate tumor growth and metastasis inhibition in vivo. RESULTS: Wound healing and transwell assays demonstrated inhibition of migration in the A549 and H226br cells 4 to 24 hours after transfection. Ectopic c-Cbl WT expression was found to reduce cell proliferation at 48 hours in A549 cells. It is important to note that A549 and H1299-Luc cells with ectopic c-Cbl WT expression demonstrated inhibition of tumor growth in vivo. A549 cells overexpressing c-Cbl WT inhibited tumor metastasis in animal models. CONCLUSIONS: To the best of the authors' knowledge, the current study is the first to demonstrate that c-Cbl WT protein overexpression inhibits tumor metastasis and tumor growth in lung cancer xenograft models. These results provide evidence that ectopic expression of c-Cbl WT protein can be potentially applied as targeted therapy for the treatment of lung cancer.

Early intervention mechanism for preventing electrocution in construction engineering.

The aim of this study is to establish an effective early intervention mechanism for construction engineering to prevent electrocution while improving labor safety and reducing the casualty risk. This study used narrative text analysis and the Haddon Matrix for data collection, and analyzed the causes from the 113 electrocution deaths among in the construction industry, the exhaustive chi-square automatic interaction detector algorithm was employed the segmentation of the correlations. Based on the theory of inventive problem solving, through IDEF0 (ICAM DEFinition) for function modeling was designed the early intervention mechanism. This study revealed the operating features related to electric shock hazards. Early intervention was introduced to reduce the relevant risks and establish safety mechanisms. The first contribution of this study is the determination of hazard correlations between operating features and conductive media, and entry point for the prevention of electrocutions. The second contribution is the suggestion of the establishment of inspection stations for electric tools, thereby ensuring that the portable power tools are safe. The final contribution is the joint application of TRIZ (Teoriya Resheniya Izobreatatelskikh Zadatch) and IDEF0, which establishing the pre-entry testing, strengthening safety mechanisms.

FoxD5 mediates anterior-posterior polarity through upstream modulator Fgf signaling during zebrafish somitogenesis.

The transcription factor FoxD5 is expressed in the paraxial mesoderm of zebrafish. However, the roles of FoxD5 in anterior pre-somitic mesoderm (PSM) during somitogenesis are unknown. We knocked down FoxD5 in embryos, which resulted in defects of the newly formed somites, including loss of the striped patterns of anterior-posterior polarity genes deltaC, notch2, notch3 and EphB2a, as well as the absence of mespa expression in S-I. Also, the expression of mespb exhibited a 'salt and pepper' pattern, indicating that FoxD5 is necessary for somite patterning in anterior PSM. Embryos were treated with SU5402, an Fgf receptor (FGFR) inhibitor, resulting in reduction of FoxD5 expression. This finding was consistent with results obtained from Tg(hsp70l:dnfgfr1-EGFP)pd1 embryos, whose dominant-negative form of FGFR1 was produced by heat-induction. Loss of FoxD5 expression was observed in the embryos injected with fgf3-/fgf8-double-morpholinos (MOs). Excessive FoxD5 mRNA could rescue the defective expression levels of mespa and mespb in fgf3-/fgf8-double morphants, suggesting that Fgf signaling acts as an upstream modulator of FoxD5 during somitogenesis. We concluded that FoxD5 is required for maintaining anterior-posterior polarity within a somite and that the striped pattern of FoxD5 in anterior PSM is mainly regulated by Fgf. An Fgf-FoxD5-Mesps signaling network is therefore proposed.

[Location, selection, and comparative anatomy of "Taixi" (KI3), "Shuiquan" (KI5), "Fuliu" (KI7), "Jiaoxin" (KI8), "Zhubin" (KI9), and "Yingu"(KI10) acupoints in rabbits].

OBJECTIVE: To investigate the. METHODS: for locating and selecting the acupoints of "Taixi" (KI3), "Shuiquan" (KI5), "Fuliu" (KI7), "Jiaoxin" (KI8), "Zhubin" (KI9), and "Yingu" (KI10) and the morphological structure of these acupoints in rabbits. MethodsAccording to the WHO and national standards for human acupoints and rabbit X-ray images, acupoint locations were marked using the anatomical landmarks on body surface in 10 New Zealand rabbits. The acupoints were dissected to compare the homologous and analogous tissue between rabbits and human body and thus correct the locations of these acupoints. Potentials were measured for the 10 New Zealand rabbits at the corrected locations of the acupoints and around the acupoints, and the final locations of these acupoints were determined by comparing the anatomical results and the data of potentials. Anatomical observation was performed after marking, and the relationship between acupuncture needle and adjacent structure was observed. RESULTS: "Taixi" was located in the ankle area, at the midpoint between the prominence of the medial malleolus and the calca-neal tendon; "Shuiquan" was located in the calcaneal area below "Taixi" in the depression anterior to the calcaneal tuberosity; "Fuliu" was located at the medial side of the calf, at 2 cun above the prominence of the medial malleolus anterior to the calcaneal tendon; "Jiaoxin" was located at the medial side of the calf, at 2 cun above the prominence of the medial malleolus and in the depression posterior to the medial border of the tibia; "Zhubin" was located at the medial side of the calf, at 5 cun above the medial malleolus on the line between "Taixi" and "Yingu"; "Yingu" was located at the medial side of the knee, at the posterior-inferior border of the semitendinosus tendon on the popliteal crease. The results of skin potentials at the acupoints suggested that "Taixi", "Shuiquan", "Fuliu", and "Zhubin" were high-reliability acupoints, "Jiaoxin" was a medium-reliability acupoint, and "Yingu" was a low-reliability acupoint. CONCLUSION: Comparative anatomy combined with imaging, surface anatomy, and electrophysiological techniques of acupoints can help with the accurate localization and selection of acupoints in experimental animals, improve the reliability of acupoint location, and enrich the comparative anatomical data of acupoints.

Overexpression of PAFAH1B1 is associated with tumor metastasis and poor survival in non-small cell lung cancer.

Our previous array-comparative genomic hybridization study showed that PAFAH1B1 gene locus was amplified in lung cancer patients, suggesting that PAFAH1B1 is a potential oncogene in lung cancer. Here, we investigate the oncogenic mechanisms of PAFAH1B1 in lung cancer. PAFAH1B1 was characterized in cell and animal models of lung cancer by in vitro migration and invasion assays and in vivo metastasis studies. The mRNA and protein expression levels of PAFAH1B1 were further determined and the prognostic effects of PAFAH1B1 overexpression in lung cancer patients were analyzed. Overexpression of PAFAH1B1 enhanced migration and invasion in lung cancer cells, whereas knockdown of PAFAH1B1 decreased cell migration and invasion, and disrupted cell microtubule organization and pericellular poly-fibronectin assemblies. In vivo tumor metastasis assay confirmed that PAFAH1B1 knockdown in lung cancer cells markedly reduced their metastasis capabilities in animals. The frequencies of overexpressed PAFAH1B1 mRNA and protein were 62.4% (63/101) and 57.4% (58/101) in lung cancer patients, respectively. The clinical correlation results showed that overexpression of PAFAH1B1 was significantly associated with late stage (mRNA: P=0.008, protein: P=0.008) and poor survival in lung adenocarcinoma (P=0.020) and male patients (P=0.049). Our results provide the first evidence that PAFAH1B1 overexpression contributes to lung tumorigenesis and poor prognosis. These effects are partly mediated through disruption of microtubule network and pericellular poly-fibronectin assembly to promote migration and invasiveness of lung cancer cells.

CBL is frequently altered in lung cancers: its relationship to mutations in MET and EGFR tyrosine kinases.

BACKGROUND: Non-small cell lung cancer (NSCLC) is a heterogeneous group of disorders with a number of genetic and proteomic alterations. c-CBL is an E3 ubiquitin ligase and adaptor molecule important in normal homeostasis and cancer. We determined the genetic variations of c-CBL, relationship to receptor tyrosine kinases (EGFR and MET), and functionality in NSCLC. METHODS AND FINDINGS: Using archival formalin-fixed paraffin embedded (FFPE) extracted genomic DNA, we show that c-CBL mutations occur in somatic fashion for lung cancers. c-CBL mutations were not mutually exclusive of MET or EGFR mutations; however they were independent of p53 and KRAS mutations. In normal/tumor pairwise analysis, there was significant loss of heterozygosity (LOH) for the c-CBL locus (22%, n = 8/37) and none of these samples revealed any mutation in the remaining copy of c-CBL. The c-CBL LOH also positively correlated with EGFR and MET mutations observed in the same samples. Using select c-CBL somatic mutations such as S80N/H94Y, Q249E and W802* (obtained from Caucasian, Taiwanese and African-American samples, respectively) transfected in NSCLC cell lines, there was increased cell viability and cell motility. CONCLUSIONS: Taking the overall mutation rate of c-CBL to be a combination as somatic missense mutation and LOH, it is clear that c-CBL is highly mutated in lung cancers and may play an essential role in lung tumorigenesis and metastasis.

Benefiting from networks by occupying central positions: an empirical study of the Taiwan health care industry.

At issue is whether network resources imply some resources available to all members in networks or available only to those occupying structurally central positions in networks. In this article, two conceptual models, the additive and interaction models of the firm, are empirically tested regarding the impact of hospital resources, network resources, and centrality on hospital performance in the Taiwan health care industry. The results demonstrate that: (1) in the additive model, hospital resources and centrality independently affect performance, whereas network resources do not; and (2) no evidence supports the interaction effect of centrality and resources on performance. Based on our findings in Taiwanese practices, the extent to which the resources are acquired externally from networks, we suggest that while adopting interorganizational strategies, hospitals should clearly identify those important resources that reside in-house and those transferred from network partners. How hospitals access resources from central positions is more important than what network resources can hospitals acquire from networks. Hospitals should improve performance by exploiting its in-house resources rather than obtaining network resources externally. In addition, hospitals should not only invest in hospital resources for better performance but should also move to central positions in networks to benefit from collaborations.