Collagenase expression in ovarian cancer cell lines

David H. Moore, Bernadette Allison, Katherine Y. Look, Gregory P. Sutton, Robert Bigsby

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Background: There are several distinct forms of the matrix metalloproteinases (MMP) that degrade collagen in the extracellular matrix. Most involved in the metastatic process are collagenases exhibiting specificity for type IV collagen, the collagen that makes up the backbone of the basement membrane. We chose to examine the expression of one type IV collagenase, MMP-9, in several ovarian cancer cell lines. Methods: Under the influence of phorbol esters such as TPA, the fibrosarcoma cell line HT1080 is known to express MMP-9. We examined the effect of TPA on MMP-9 expression with Northern analysis, and on MMP-9 collagenase activity with polyacrylamide gel electrophoresis-zymography, in HT1080 cells (positive control) and in six ovarian cancer cell lines: Hey, HeyA8, CAOV3, PA-1, SKOV3, and OVCAR3. Cells were grown to 70% confluence and the growth medium was changed to a serum- free medium for 24 hr. Cells were then treated with 12 ng/mL TPA or its vehicle for an additional 24 hr. Medium was collected for PAGE-zymographic analysis. The cells were lysed and their RNA collected for Northern analysis. Results: PA-1, a teratocarcinoma, expressed more than 10-fold the amount of RNA for MMP-9 than the other lines following TPA treatment. Both Northern analysis and zymography showed that four of the six ovarian cell lines responded to TPA with increased collagenase expression and activity. The other two cell lines showed no MMP-9 activity either before or after TPA treatment. Conclusions: These results suggest that ovarian cancer cell lines express collagenase and that this expression may be regulated by phorbol esters which activate the protein kinase C pathway.

Original languageEnglish
Pages (from-to)78-82
Number of pages5
JournalGynecologic Oncology
Volume65
Issue number1
DOIs
StatePublished - Apr 1997

Fingerprint

Matrix Metalloproteinase 9
Collagenases
Ovarian Neoplasms
Cell Line
Phorbol Esters
Collagen
RNA
Teratocarcinoma
Collagen Type IV
Fibrosarcoma
Serum-Free Culture Media
Matrix Metalloproteinases
Basement Membrane
Protein Kinase C
Extracellular Matrix
Polyacrylamide Gel Electrophoresis
Growth

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Oncology

Cite this

Collagenase expression in ovarian cancer cell lines. / Moore, David H.; Allison, Bernadette; Look, Katherine Y.; Sutton, Gregory P.; Bigsby, Robert.

In: Gynecologic Oncology, Vol. 65, No. 1, 04.1997, p. 78-82.

Research output: Contribution to journalArticle

Moore, DH, Allison, B, Look, KY, Sutton, GP & Bigsby, R 1997, 'Collagenase expression in ovarian cancer cell lines', Gynecologic Oncology, vol. 65, no. 1, pp. 78-82. https://doi.org/10.1006/gyno.1997.4628
Moore, David H. ; Allison, Bernadette ; Look, Katherine Y. ; Sutton, Gregory P. ; Bigsby, Robert. / Collagenase expression in ovarian cancer cell lines. In: Gynecologic Oncology. 1997 ; Vol. 65, No. 1. pp. 78-82.
@article{04f841e376804d64a08905f31d954d1b,
title = "Collagenase expression in ovarian cancer cell lines",
abstract = "Background: There are several distinct forms of the matrix metalloproteinases (MMP) that degrade collagen in the extracellular matrix. Most involved in the metastatic process are collagenases exhibiting specificity for type IV collagen, the collagen that makes up the backbone of the basement membrane. We chose to examine the expression of one type IV collagenase, MMP-9, in several ovarian cancer cell lines. Methods: Under the influence of phorbol esters such as TPA, the fibrosarcoma cell line HT1080 is known to express MMP-9. We examined the effect of TPA on MMP-9 expression with Northern analysis, and on MMP-9 collagenase activity with polyacrylamide gel electrophoresis-zymography, in HT1080 cells (positive control) and in six ovarian cancer cell lines: Hey, HeyA8, CAOV3, PA-1, SKOV3, and OVCAR3. Cells were grown to 70{\%} confluence and the growth medium was changed to a serum- free medium for 24 hr. Cells were then treated with 12 ng/mL TPA or its vehicle for an additional 24 hr. Medium was collected for PAGE-zymographic analysis. The cells were lysed and their RNA collected for Northern analysis. Results: PA-1, a teratocarcinoma, expressed more than 10-fold the amount of RNA for MMP-9 than the other lines following TPA treatment. Both Northern analysis and zymography showed that four of the six ovarian cell lines responded to TPA with increased collagenase expression and activity. The other two cell lines showed no MMP-9 activity either before or after TPA treatment. Conclusions: These results suggest that ovarian cancer cell lines express collagenase and that this expression may be regulated by phorbol esters which activate the protein kinase C pathway.",
author = "Moore, {David H.} and Bernadette Allison and Look, {Katherine Y.} and Sutton, {Gregory P.} and Robert Bigsby",
year = "1997",
month = "4",
doi = "10.1006/gyno.1997.4628",
language = "English",
volume = "65",
pages = "78--82",
journal = "Gynecologic Oncology",
issn = "0090-8258",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Collagenase expression in ovarian cancer cell lines

AU - Moore, David H.

AU - Allison, Bernadette

AU - Look, Katherine Y.

AU - Sutton, Gregory P.

AU - Bigsby, Robert

PY - 1997/4

Y1 - 1997/4

N2 - Background: There are several distinct forms of the matrix metalloproteinases (MMP) that degrade collagen in the extracellular matrix. Most involved in the metastatic process are collagenases exhibiting specificity for type IV collagen, the collagen that makes up the backbone of the basement membrane. We chose to examine the expression of one type IV collagenase, MMP-9, in several ovarian cancer cell lines. Methods: Under the influence of phorbol esters such as TPA, the fibrosarcoma cell line HT1080 is known to express MMP-9. We examined the effect of TPA on MMP-9 expression with Northern analysis, and on MMP-9 collagenase activity with polyacrylamide gel electrophoresis-zymography, in HT1080 cells (positive control) and in six ovarian cancer cell lines: Hey, HeyA8, CAOV3, PA-1, SKOV3, and OVCAR3. Cells were grown to 70% confluence and the growth medium was changed to a serum- free medium for 24 hr. Cells were then treated with 12 ng/mL TPA or its vehicle for an additional 24 hr. Medium was collected for PAGE-zymographic analysis. The cells were lysed and their RNA collected for Northern analysis. Results: PA-1, a teratocarcinoma, expressed more than 10-fold the amount of RNA for MMP-9 than the other lines following TPA treatment. Both Northern analysis and zymography showed that four of the six ovarian cell lines responded to TPA with increased collagenase expression and activity. The other two cell lines showed no MMP-9 activity either before or after TPA treatment. Conclusions: These results suggest that ovarian cancer cell lines express collagenase and that this expression may be regulated by phorbol esters which activate the protein kinase C pathway.

AB - Background: There are several distinct forms of the matrix metalloproteinases (MMP) that degrade collagen in the extracellular matrix. Most involved in the metastatic process are collagenases exhibiting specificity for type IV collagen, the collagen that makes up the backbone of the basement membrane. We chose to examine the expression of one type IV collagenase, MMP-9, in several ovarian cancer cell lines. Methods: Under the influence of phorbol esters such as TPA, the fibrosarcoma cell line HT1080 is known to express MMP-9. We examined the effect of TPA on MMP-9 expression with Northern analysis, and on MMP-9 collagenase activity with polyacrylamide gel electrophoresis-zymography, in HT1080 cells (positive control) and in six ovarian cancer cell lines: Hey, HeyA8, CAOV3, PA-1, SKOV3, and OVCAR3. Cells were grown to 70% confluence and the growth medium was changed to a serum- free medium for 24 hr. Cells were then treated with 12 ng/mL TPA or its vehicle for an additional 24 hr. Medium was collected for PAGE-zymographic analysis. The cells were lysed and their RNA collected for Northern analysis. Results: PA-1, a teratocarcinoma, expressed more than 10-fold the amount of RNA for MMP-9 than the other lines following TPA treatment. Both Northern analysis and zymography showed that four of the six ovarian cell lines responded to TPA with increased collagenase expression and activity. The other two cell lines showed no MMP-9 activity either before or after TPA treatment. Conclusions: These results suggest that ovarian cancer cell lines express collagenase and that this expression may be regulated by phorbol esters which activate the protein kinase C pathway.

UR - http://www.scopus.com/inward/record.url?scp=0031128265&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031128265&partnerID=8YFLogxK

U2 - 10.1006/gyno.1997.4628

DO - 10.1006/gyno.1997.4628

M3 - Article

VL - 65

SP - 78

EP - 82

JO - Gynecologic Oncology

JF - Gynecologic Oncology

SN - 0090-8258

IS - 1

ER -