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The interrelationship between DRIM gene expression and cytogenetic and phenotypic characteristics in human breast tumor cell lines.

Goodison S, Viars C, Grazzini M, Urquidi V - BMC Genomics (2003)

Bottom Line: DRIM encodes a protein comprising 2785 amino acids with significant homology to a protein in yeast and C. elegans, but little else is currently known about its function or pattern of expression.We confirmed higher expression of DRIM mRNA in the non-metastatic MDA-MB-435 clone NM2C5, relative to its metastatic counterpart M4A4, but this appeared to be due to the presence of an extra copy of the DRIM gene in the cell line's genome.The other non-metastatic cell lines in the series (T47D MCF-7, SK-BR-3 and ZR-75-1) contained either 3 or 4 chromosomal copies of DRIM gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: UCSD Cancer Center and Department of Pathology, University of California, San Diego, La Jolla, CA, USA. sgoodison@ucsd.edu

ABSTRACT

Background: In order to facilitate the identification of genes involved in the metastatic phenotype we have previously developed a pair of cell lines from the human breast carcinoma cell line MDA-MB-435, which have diametrically opposite metastatic potential in athymic mice. Differential display analysis of this model previously identified a novel gene, DRIM (down regulated in metastasis), the decreased expression of which correlated with metastatic capability. DRIM encodes a protein comprising 2785 amino acids with significant homology to a protein in yeast and C. elegans, but little else is currently known about its function or pattern of expression. In a detailed analysis of the DRIM gene locus we quantitatively evaluated gene dosage and the expression of DRIM transcripts in a panel of breast cell lines of known metastatic phenotype.

Results: Fluorescent in situ hybridization (FISH) analyses mapped a single DRIM gene locus to human chromosome 12q23~24, a region of conserved synteny to mouse chromosome 10. We confirmed higher expression of DRIM mRNA in the non-metastatic MDA-MB-435 clone NM2C5, relative to its metastatic counterpart M4A4, but this appeared to be due to the presence of an extra copy of the DRIM gene in the cell line's genome. The other non-metastatic cell lines in the series (T47D MCF-7, SK-BR-3 and ZR-75-1) contained either 3 or 4 chromosomal copies of DRIM gene. However, the expression level of DRIM mRNA in M4A4 was found to be 2-4 fold higher than in unrelated breast cells of non-metastatic phenotype.

Conclusions: Whilst DRIM expression is decreased in metastatic M4A4 cells relative to its non-metastatic isogenic counterpart, neither DRIM gene dosage nor DRIM mRNA levels correlated with metastatic propensity in a series of human breast tumor cell lines examined. Collectively, these findings indicate that the expression pattern of the DRIM gene in relation to the pathogenesis of breast tumor metastasis is more complex than previously recognized.

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(A) genomic chromosomal localization of the human DRIM by fluorescence in situ hybridization. Metaphase spread counterstained with DAPI and hybridized with a DRIM genomic DNA probe (red) demonstrating DRIM-specific signals on the long arm of chromosome 12. (B) additional images of chromosome 12 showing DRIM localization at 12q23~24. (a) counterstained with DAPI (DRIM-specific signal in red)., (b) an image of reverse-DAPI banding., and (c) an R-banded chromosome (DRIM-specific signal in yellow).
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Figure 1: (A) genomic chromosomal localization of the human DRIM by fluorescence in situ hybridization. Metaphase spread counterstained with DAPI and hybridized with a DRIM genomic DNA probe (red) demonstrating DRIM-specific signals on the long arm of chromosome 12. (B) additional images of chromosome 12 showing DRIM localization at 12q23~24. (a) counterstained with DAPI (DRIM-specific signal in red)., (b) an image of reverse-DAPI banding., and (c) an R-banded chromosome (DRIM-specific signal in yellow).

Mentions: The chromosomal localization of the human DRIM locus was clarified and refined by FISH using metaphase spreads obtained from normal human peripheral blood leukocytes, and a combination probe composed of two human DRIM genomic clones. Signals were considered specific only if they were detected on both chromatids of a single chromosome. Applying this criteria, specific DRIM signals were detected in 18 of 20 metaphases examined (90%), and in each case, the hybridization signals were located on the long arm of chromosome 12 (Figure 1). A combination of DAPI and R-banding of chromosomes enabled the assignment of the DRIM locus to 12q23~24. The localization of DRIM on chromosome 12 was further confirmed by two-color FISH using both the DRIM probe and a chromosome 12-specific paint (data not shown). No DRIM signals were detected on any other chromosome. This chromosomal location is consistent with information available from the NCBI Map View database which places DRIM at 12q23 between markers D12S346-D12S78. The location is also consistent with the localization of the 156 bp STS G3 1980 (Sanger Center's GB4 radiation hybrid RH65919) which shows 100% homology to DRIM and has been placed 689.3 cR from the top of chromosome 12 (Figure 2).


The interrelationship between DRIM gene expression and cytogenetic and phenotypic characteristics in human breast tumor cell lines.

Goodison S, Viars C, Grazzini M, Urquidi V - BMC Genomics (2003)

(A) genomic chromosomal localization of the human DRIM by fluorescence in situ hybridization. Metaphase spread counterstained with DAPI and hybridized with a DRIM genomic DNA probe (red) demonstrating DRIM-specific signals on the long arm of chromosome 12. (B) additional images of chromosome 12 showing DRIM localization at 12q23~24. (a) counterstained with DAPI (DRIM-specific signal in red)., (b) an image of reverse-DAPI banding., and (c) an R-banded chromosome (DRIM-specific signal in yellow).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC222913&req=5

Figure 1: (A) genomic chromosomal localization of the human DRIM by fluorescence in situ hybridization. Metaphase spread counterstained with DAPI and hybridized with a DRIM genomic DNA probe (red) demonstrating DRIM-specific signals on the long arm of chromosome 12. (B) additional images of chromosome 12 showing DRIM localization at 12q23~24. (a) counterstained with DAPI (DRIM-specific signal in red)., (b) an image of reverse-DAPI banding., and (c) an R-banded chromosome (DRIM-specific signal in yellow).
Mentions: The chromosomal localization of the human DRIM locus was clarified and refined by FISH using metaphase spreads obtained from normal human peripheral blood leukocytes, and a combination probe composed of two human DRIM genomic clones. Signals were considered specific only if they were detected on both chromatids of a single chromosome. Applying this criteria, specific DRIM signals were detected in 18 of 20 metaphases examined (90%), and in each case, the hybridization signals were located on the long arm of chromosome 12 (Figure 1). A combination of DAPI and R-banding of chromosomes enabled the assignment of the DRIM locus to 12q23~24. The localization of DRIM on chromosome 12 was further confirmed by two-color FISH using both the DRIM probe and a chromosome 12-specific paint (data not shown). No DRIM signals were detected on any other chromosome. This chromosomal location is consistent with information available from the NCBI Map View database which places DRIM at 12q23 between markers D12S346-D12S78. The location is also consistent with the localization of the 156 bp STS G3 1980 (Sanger Center's GB4 radiation hybrid RH65919) which shows 100% homology to DRIM and has been placed 689.3 cR from the top of chromosome 12 (Figure 2).

Bottom Line: DRIM encodes a protein comprising 2785 amino acids with significant homology to a protein in yeast and C. elegans, but little else is currently known about its function or pattern of expression.We confirmed higher expression of DRIM mRNA in the non-metastatic MDA-MB-435 clone NM2C5, relative to its metastatic counterpart M4A4, but this appeared to be due to the presence of an extra copy of the DRIM gene in the cell line's genome.The other non-metastatic cell lines in the series (T47D MCF-7, SK-BR-3 and ZR-75-1) contained either 3 or 4 chromosomal copies of DRIM gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: UCSD Cancer Center and Department of Pathology, University of California, San Diego, La Jolla, CA, USA. sgoodison@ucsd.edu

ABSTRACT

Background: In order to facilitate the identification of genes involved in the metastatic phenotype we have previously developed a pair of cell lines from the human breast carcinoma cell line MDA-MB-435, which have diametrically opposite metastatic potential in athymic mice. Differential display analysis of this model previously identified a novel gene, DRIM (down regulated in metastasis), the decreased expression of which correlated with metastatic capability. DRIM encodes a protein comprising 2785 amino acids with significant homology to a protein in yeast and C. elegans, but little else is currently known about its function or pattern of expression. In a detailed analysis of the DRIM gene locus we quantitatively evaluated gene dosage and the expression of DRIM transcripts in a panel of breast cell lines of known metastatic phenotype.

Results: Fluorescent in situ hybridization (FISH) analyses mapped a single DRIM gene locus to human chromosome 12q23~24, a region of conserved synteny to mouse chromosome 10. We confirmed higher expression of DRIM mRNA in the non-metastatic MDA-MB-435 clone NM2C5, relative to its metastatic counterpart M4A4, but this appeared to be due to the presence of an extra copy of the DRIM gene in the cell line's genome. The other non-metastatic cell lines in the series (T47D MCF-7, SK-BR-3 and ZR-75-1) contained either 3 or 4 chromosomal copies of DRIM gene. However, the expression level of DRIM mRNA in M4A4 was found to be 2-4 fold higher than in unrelated breast cells of non-metastatic phenotype.

Conclusions: Whilst DRIM expression is decreased in metastatic M4A4 cells relative to its non-metastatic isogenic counterpart, neither DRIM gene dosage nor DRIM mRNA levels correlated with metastatic propensity in a series of human breast tumor cell lines examined. Collectively, these findings indicate that the expression pattern of the DRIM gene in relation to the pathogenesis of breast tumor metastasis is more complex than previously recognized.

Show MeSH
Related in: MedlinePlus