Genomic profiling of myeloid sarcoma by array comparative genomic hybridization.

Myeloid sarcoma (MS) is a tumor mass of myeloblasts or immature myeloid cells occurring in an extramedullary site. In this study, seven cases of MS [stomach (1), testis (1), skin (2), and lymph node (3)] and 3 synchronous and 1 follow-up bone marrow (BM) samples were studied for genomic abnormalities using array comparative genomic hybridization (array-CGH). Array-CGH construction used approximately 5,400 bacterial artificial chromosome clones from the RPCI-11 library, spanning the human genome.

EVI5 is a novel centrosomal protein that binds to alpha- and gamma-tubulin.

The human EVI5 protein carries a TBC domain indicative of Rab GTPase activating protein (GAP) activity, and an extensive coiled-coil motif in the C-terminal region. EVI5 is ubiquitously expressed in adult, fetal, and cancer tissues and exists as two mRNA species resulting from differential use of polyadenylation signals. Western blot analysis suggests that different molecular weight protein species are probably generated by posttranslational modification.

Differential expression of the LGI and SLIT families of genes in human cancer cells.

The LGI and SLIT genes have a distinctive leucine-rich repeat motif in the N-terminal end of the protein which is indicative of either receptor function or an interaction with the extracellular matrix. Members of the LGI and SLIT family of genes have been implicated in specific cancers and have been suggested to have a restricted pattern of expression in normal cells. To investigate the extent and distribution of the expression of these genes in cancer cells we have analyzed their expression levels in a range of tumor cell types.

Mass spectroscopy identifies the splicing-associated proteins, PSF, hnRNP H3, hnRNP A2/B1, and TLS/FUS as interacting partners of the ZNF198 protein associated with rearrangement in myeloproliferative disease.

ZNF198 is fused with FGFR1 in an atypical myeloproliferative disease that results in constitutive activation of the kinase domain and mislocalization to the cytoplasm. We have used immunoprecipitation of a GFP-tagged ZNF198 combined with MALDI-TOF mass spectroscopy to identify interacting proteins. P splicing factor (PSF) was identified as one of the proteins and this interaction was confirmed by Western blotting.

Identification of consistent novel submegabase deletions in low-grade oligodendrogliomas using array-based comparative genomic hybridization.

We have analyzed 18 low-grade gliomas using array comparative genomic hybridization (aCGH) with an average resolution of <500 kb. Because the majority of these tumors showed loss of chromosome arms 1p and 19q, we used custom statistical approaches to define submegabase hemizygous losses throughout the genome that correlated with 19q loss. As a result of this analysis, we have identified a approximately 550-kb region in 11q13 and a approximately 300-kb region in 13q12 that showed hemizygous deletion in virtually all the tumors analyzed regardless of their 1p/19q status.

WAVE3 promotes cell motility and invasion through the regulation of MMP-1, MMP-3, and MMP-9 expression.

WAVE3 is a member of the WASP/WAVE family of proteins, which play a critical role in the regulation of actin polymerization, cytoskeleton organization, and cell motility. We show here that knockdown of the WAVE3 protein, using RNA interference in MDA-MB-231 cells, decreases phospho-p38 MAPK levels, but not those of phospho-AKT, phospho-ERK, or phospho-JNK. Knockdown of WAVE3 expression also inhibited the expression levels of MMP-1, MMP-3, and MMP-9, but not MMP-2.

Molecular characterization of a consistent 4.5-megabase deletion at 4q28 in prostate cancer cells.

Spectral karyotyping of prostate cell lines LNCaP, DU145, PC3, and 22RV demonstrated structural chromosome rearrangements involving the distal long arm of chromosome 4. In all but 22RV, these are nonreciprocal translocations between chromosomes 4 and 10. In 22RV, an apparently reciprocal t(2q;4q) is seen.

WAVE3-mediated cell migration and lamellipodia formation are regulated downstream of phosphatidylinositol 3-kinase.

WAVE3 is a member of the WASP/WAVE family of protein effectors of actin reorganization and cell movement. The precise role of WAVE3 in cell migration and its regulation, however, have not been elucidated. Here we show that endogenous WAVE3 was found to be concentrated in the lamellipodia at the leading edge of migrating MDA-MB-231 cells.

Analysis of the RB1 gene in children with retinoblastoma having residential connections to West Cumbria, England.

Six of eight cases of retinoblastoma previously identified as having a residential association with West Cumbria, England, in which the Sellafield nuclear installation is situated, were examined for the presence of a constitutional RB1 mutation. No mutations were detected, thus providing strong evidence against an environmental or occupational genotoxic effect causing germline mutations in the parents of these children.

The relationship between maternal and child mental health in Mexican immigrant families.

Depression is a leading cause of disability in the United States, with Mexican immigrant women reporting depression rates higher than the national average. The purposes of this study were to describe mental health symptoms in a sample (n = 182) of Mexican immigrant mothers and their relationships to child mental health, family functioning, and acculturation. Over one third of the mothers reported depression and anxiety symptoms above standardized cutoffs while 31% of the children scored in the depressed range.

High throughput determination of gains and losses of genetic material using high resolution BAC arrays and comparative genomic hybridization.

Chromosome analysis has been a cornerstone both for the identification of genetic defects that predispose to a variety of genetic syndromes as well as for the analysis of cancer progression. The relatively low resolution of metaphase chromosomes, however, only allows characterization of major genetic events which are defined at the megabase level. The development of the human genome-wide bacterial artificial chromosome (BACs) libraries which were used as templates for the human genome project made it possible to design microarrays containing these BACs which can theoretically span the genome uninterrupted.

Obtaining DNA from a geographically dispersed cohort of current and former smokers: use of mail-based mouthwash collection and monetary incentives.

The feasibility of collecting DNA through the mail from a cohort of current and former smokers was assessed. Also examined was whether monetary incentives would increase response rates. A random sample of 300 subjects, stratified by 20 U.

Application of bacterial artificial chromosome array-based comparative genomic hybridization and spectral karyotyping to the analysis of glioblastoma multiforme.

Identification of genetic losses and gains is valuable in analysis of brain tumors. Locus-by-locus analyses have revealed correlations between prognosis and response to chemotherapy and loss or gain of specific genes and loci. These approaches are labor intensive and do not provide a global view of the genetic changes within the tumor cells.

LGI1, a putative tumor metastasis suppressor gene, controls in vitro invasiveness and expression of matrix metalloproteinases in glioma cells through the ERK1/2 pathway.

Gliomas take a number of different genetic routes in the progression to glioblastoma multiforme, a highly invasive variant that is mostly unresponsive to current therapies. Gliomas express elevated levels of matrix metalloproteinases (MMPs), which have been implicated in the control of proliferation and invasion as well as neovascularization. Progressive loss of LGI1 expression has been associated with the development of high grade gliomas.

Characterization of the 1p/19q chromosomal loss in oligodendrogliomas using comparative genomic hybridization arrays (CGHa).

Loss of genetic material from the short arm of chromosome 1 and the long arm of chromosome 19 in anaplastic oligodendrogliomas has been shown to predict responsiveness to chemotherapy. Currently, the most common approach used to detect this loss of 1p/19q material employs microsatellite/FISH analysis using markers along the length of these chromosome arms. This analysis is highly focused and carried out on a locus-by-locus basis and gives no indication of the extent of other genetic changes occurring in the tumor cells, which may be important in future studies to explore genetic heterogeneity in the response to treatment.

Integrating research into teaching public health nursing.

Integration of research into teaching provides an environment for students to not only learn how research is conducted but also experience how research contributes to improving practice and client outcomes. Integrating research into teaching is important because of the need to build evidence for public health nursing practice. This article describes an innovative approach by faculty to integrate research into teaching undergraduate and graduate public health nursing courses.

CLCA2 tumour suppressor gene in 1p31 is epigenetically regulated in breast cancer.

The calcium-activated chloride channel gene family is clustered in the 1p31 region, which is frequently deleted in sporadic breast cancer. Recent studies have indicated the association of the second member of this gene family (CLCA2) with the development of breast cancer and metastasis. We have now shown the absence of expression of CLCA2 in several breast cancer tumours and cell lines, which confirms the results from other reports.

Identification and characterisation of constitutional chromosome abnormalities using arrays of bacterial artificial chromosomes.

Constitutional chromosome deletions and duplications frequently predispose to the development of a wide variety of cancers. We have developed a microarray of 6000 bacterial artificial chromosomes for array-based comparative genomic hybridisation, which provides an average resolution of 750 kb across the human genome. Using these arrays, subtle gains and losses of chromosome regions can be detected in constitutional cells, following a single overnight hybridisation.

Manipulation of nonsense mediated decay identifies gene mutations in colon cancer Cells with microsatellite instability.

Cancer cells showing microsatellite instability (MSI) demonstrate a high frequency of acquired frameshift mutations that result in the generation of nonsense mutations. RNA transcripts carrying these nonsense mutations are usually targeted for degradation through the nonsense mediated decay (NMD) pathway. Blocking this pathway with drugs such as emitine, results in the 'stabilization' of these mutant transcripts, which can now be detected on cDNA arrays.

A role for p300/CREB binding protein genes in promoting cancer progression in colon cancer cell lines with microsatellite instability.

Our manipulation of the nonsense-mediated decay pathway in microsatellite unstable colon cancer cell lines identified the p300 gene as a potential tumor suppressor in this subtype of cancer. Here, we have demonstrated that not only the p300 gene but also the highly homologous cAMP-response element-binding protein (CREB) binding protein (CBP) gene together are mutated in >85% of microsatellite instability (MSI)+ colon cancer cell lines. A limited survey of primary tumors with MSI+ shows that p300 is also frequently mutated in these cancers, demonstrating that these mutations are not consequences of in vitro growth.

High-resolution analysis of genetic events in cancer cells using bacterial artificial chromosome arrays and comparative genome hybridization.

Chromosome analysis of cancer cells has been one of the primary means of identifying key genetic events in the development of cancer. The relatively low resolution of metaphase chromosomes, however, only allows characterization of major genetic events that are defined at the megabase level. The development of the human genome-wide bacterial artificial chromosome (BAC) libraries that were used as templates for the human genome project made it possible to design microarrays containing these BACs that can theoretically span the genome uninterrupted.

Genomic organization and expression profile of the human and mouse WAVE gene family.

The WAVE gene family, which contains three members, has been shown to play a major role in the actin polymerization and cytoskeleton organization processes. We have identified the WAVE3 gene from Chromosome (Chr) 13q12, as being involved in one of the breakpoints of a t(1:13)(q21:q12) reciprocal translocation, in a patient with ganglioneuroblastoma (Sossey-Alaoui et al. 2002; Oncogene 21: 5967-5974).

Suppression of the cell proliferation and invasion phenotypes in glioma cells by the LGI1 gene.

The leucine-rich, glioma-inactivated (LGI1) gene, located in 10q24, was originally identified because it was interrupted and inactivated by a reciprocal chromosome translocation in the T98G glioma cell line. Loss of LGI1 expression in high-grade brain tumors is correlated with the frequent loss of chromosome 10 during progression of gliomas. To investigate whether this gene can suppress the malignant phenotype in glioma cells, we introduced the LGI1 gene into cells that do (U87) and do not (T98G and A172) express LGI1 endogenously.