Variability in melanoma metalloproteinase expression profiling

Orsolya Giricz, Janelle L. Lauer, Gregg B. Fields

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The proteolytic activities of a disintegrin and metalloproteinase (ADAM); a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), and matrix metalloproteinase (MMP) families play important roles in normal and multiple pathological conditions. These metalloproteases have potential roles in the degradation of the ECM and in the processing of bioactive molecules. In the present study, RNA was isolated from multiple normal fibroblast and metastatic melanoma cell lines, as well as the isogenic normal tissue and tumor samples, and the gene expression levels of six ADAMs, eight MMPs, and four ADAMTSs were analyzed by real-time PCR. This approach allowed for detected changes in mRNA expression of the individual metalloproteinase genes to be compared between normal and metastatic states and also between tissue and cultured cells. Increased gene expression of several ADAM and MMP family members (MMP1, MMP8, MMP15, and ADAM15) occurred in melanoma tissue and was replicated in tissue cultures. In general, the level of ADAM and MMP mRNA expression was several-fold higher in cultured cells compared with the isogenic tissue from which they were derived. Passage-dependent expression patterns were observed for MMP8 and MMP9 in in-house-derived metastatic melanoma cell lines. This reiterates earlier suggestions that experiments using cells that have been maintained in culture should be interpreted with great care.

Original languageEnglish (US)
Pages (from-to)194-204
Number of pages11
JournalJournal of Biomolecular Techniques
Volume21
Issue number4
StatePublished - Dec 2010

Fingerprint

Metalloproteases
Disintegrins
Melanoma
Matrix Metalloproteinases
Cultured Cells
Thrombospondins
Gene Expression
Cell Line
Messenger RNA
Real-Time Polymerase Chain Reaction
Fibroblasts
RNA
Genes
Neoplasms

Keywords

  • ADAM
  • MMP
  • Tissue

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Variability in melanoma metalloproteinase expression profiling. / Giricz, Orsolya; Lauer, Janelle L.; Fields, Gregg B.

In: Journal of Biomolecular Techniques, Vol. 21, No. 4, 12.2010, p. 194-204.

Research output: Contribution to journalArticle

Giricz, Orsolya ; Lauer, Janelle L. ; Fields, Gregg B. / Variability in melanoma metalloproteinase expression profiling. In: Journal of Biomolecular Techniques. 2010 ; Vol. 21, No. 4. pp. 194-204.
@article{46cf71615be54587acdf1110f750daf5,
title = "Variability in melanoma metalloproteinase expression profiling",
abstract = "The proteolytic activities of a disintegrin and metalloproteinase (ADAM); a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), and matrix metalloproteinase (MMP) families play important roles in normal and multiple pathological conditions. These metalloproteases have potential roles in the degradation of the ECM and in the processing of bioactive molecules. In the present study, RNA was isolated from multiple normal fibroblast and metastatic melanoma cell lines, as well as the isogenic normal tissue and tumor samples, and the gene expression levels of six ADAMs, eight MMPs, and four ADAMTSs were analyzed by real-time PCR. This approach allowed for detected changes in mRNA expression of the individual metalloproteinase genes to be compared between normal and metastatic states and also between tissue and cultured cells. Increased gene expression of several ADAM and MMP family members (MMP1, MMP8, MMP15, and ADAM15) occurred in melanoma tissue and was replicated in tissue cultures. In general, the level of ADAM and MMP mRNA expression was several-fold higher in cultured cells compared with the isogenic tissue from which they were derived. Passage-dependent expression patterns were observed for MMP8 and MMP9 in in-house-derived metastatic melanoma cell lines. This reiterates earlier suggestions that experiments using cells that have been maintained in culture should be interpreted with great care.",
keywords = "ADAM, MMP, Tissue",
author = "Orsolya Giricz and Lauer, {Janelle L.} and Fields, {Gregg B.}",
year = "2010",
month = "12",
language = "English (US)",
volume = "21",
pages = "194--204",
journal = "Journal of Biomolecular Techniques",
issn = "1524-0215",
publisher = "Association of Biomolecular Resource Facilities",
number = "4",

}

TY - JOUR

T1 - Variability in melanoma metalloproteinase expression profiling

AU - Giricz, Orsolya

AU - Lauer, Janelle L.

AU - Fields, Gregg B.

PY - 2010/12

Y1 - 2010/12

N2 - The proteolytic activities of a disintegrin and metalloproteinase (ADAM); a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), and matrix metalloproteinase (MMP) families play important roles in normal and multiple pathological conditions. These metalloproteases have potential roles in the degradation of the ECM and in the processing of bioactive molecules. In the present study, RNA was isolated from multiple normal fibroblast and metastatic melanoma cell lines, as well as the isogenic normal tissue and tumor samples, and the gene expression levels of six ADAMs, eight MMPs, and four ADAMTSs were analyzed by real-time PCR. This approach allowed for detected changes in mRNA expression of the individual metalloproteinase genes to be compared between normal and metastatic states and also between tissue and cultured cells. Increased gene expression of several ADAM and MMP family members (MMP1, MMP8, MMP15, and ADAM15) occurred in melanoma tissue and was replicated in tissue cultures. In general, the level of ADAM and MMP mRNA expression was several-fold higher in cultured cells compared with the isogenic tissue from which they were derived. Passage-dependent expression patterns were observed for MMP8 and MMP9 in in-house-derived metastatic melanoma cell lines. This reiterates earlier suggestions that experiments using cells that have been maintained in culture should be interpreted with great care.

AB - The proteolytic activities of a disintegrin and metalloproteinase (ADAM); a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), and matrix metalloproteinase (MMP) families play important roles in normal and multiple pathological conditions. These metalloproteases have potential roles in the degradation of the ECM and in the processing of bioactive molecules. In the present study, RNA was isolated from multiple normal fibroblast and metastatic melanoma cell lines, as well as the isogenic normal tissue and tumor samples, and the gene expression levels of six ADAMs, eight MMPs, and four ADAMTSs were analyzed by real-time PCR. This approach allowed for detected changes in mRNA expression of the individual metalloproteinase genes to be compared between normal and metastatic states and also between tissue and cultured cells. Increased gene expression of several ADAM and MMP family members (MMP1, MMP8, MMP15, and ADAM15) occurred in melanoma tissue and was replicated in tissue cultures. In general, the level of ADAM and MMP mRNA expression was several-fold higher in cultured cells compared with the isogenic tissue from which they were derived. Passage-dependent expression patterns were observed for MMP8 and MMP9 in in-house-derived metastatic melanoma cell lines. This reiterates earlier suggestions that experiments using cells that have been maintained in culture should be interpreted with great care.

KW - ADAM

KW - MMP

KW - Tissue

UR - http://www.scopus.com/inward/record.url?scp=78650143782&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78650143782&partnerID=8YFLogxK

M3 - Article

VL - 21

SP - 194

EP - 204

JO - Journal of Biomolecular Techniques

JF - Journal of Biomolecular Techniques

SN - 1524-0215

IS - 4

ER -