Higher expression and activity of metalloproteinases in human cervical carcinoma cell lines is associated with HPV presence

Matrix metalloproteinases (MMPs) MMP–2, MMP–9, and MT1-MMP are required for basement membrane degradation in cervical carcinoma. We evaluated the expression and activity of MMPs and their inhibitors RECK and TIMP-2 in 3 human invasive cervical carcinoma cell lines. Two HPV16-positive cell lines (SiHa and CaSki) and an HPV-negative cell line (C33A) were cultured either onto a type-I collagen gel, Matrigel™, or plastic, to recreate their three-dimensional growth environment and evaluate the expression of these genes using quantitative real-time PCR. We also analyzed the gelatinolytic activity of MMP-2 and MMP–9 by zymography. We found that HPV (human papillomavirus)-positive cell lines express higher levels of MMP-2, MT1-MMP, and TIMP-2 than the HPV negative cell line. In addition, MMP-9 was expressed at very low levels in both HPV-negative and HPV-positive cell lines. We also observed that the expression of the RECK gene is higher in CaSki cells, being associated with higher pro-MMP-2 activity. Furthermore, Matrigel™ substrate influences MMP-2 expression in both SiHa and CaSki cells. On the other hand, we found that type-I collagen gel, but not Matrigel™, can enhance pro-MMP-2 activity in all cell lines. Our results suggest that the presence of HPV is related to increased expression of MMP-2, MT1-MMP, and TIMP-2, and that pro-MMP-2 activity is higher in HPV-positive than in HPV-negative cells.

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The status of the p53 gene in human papilloma virus positive or negative cervical carcinoma cell lines

Carcinogenesis ◽

10.1093/carcin/13.7.1273 ◽

1992 ◽

Vol 13 (7) ◽

pp. 1273-1275 ◽

Cited By ~ 26

Author(s):

Shiv Srivastava ◽

Yue A. Tong ◽

Krishnakumar Devadas ◽

Zhi-Qiang Zou ◽

Yawen Chen ◽

...

Keyword(s):

Human Papilloma Virus ◽

Cell Lines ◽

Cervical Carcinoma ◽

Carcinoma Cell ◽

P53 Gene ◽

Papilloma Virus ◽

Carcinoma Cell Lines ◽

The Status

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Structure of the two promoters of the human lck gene: differential accumulation of two classes of lck transcripts in T cells

Molecular and Cellular Biology ◽

10.1128/mcb.9.5.2173-2180.1989 ◽

1989 ◽

Vol 9 (5) ◽

pp. 2173-2180

Author(s):

T Takadera ◽

S Leung ◽

A Gernone ◽

Y Koga ◽

Y Takihara ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cell Lines ◽

Carcinoma Cell ◽

Human Peripheral Blood ◽

Specific Gene ◽

Type I ◽

Type Ii ◽

Carcinoma Cell Lines ◽

Human T Cell

The human T-cell- or lymphocyte-specific gene, lck, encodes a tyrosine kinase and is a member of the src family. In this report we demonstrate that there are two classes of human lck transcripts (types I and II), containing different 5'-untranslated regions, which are expressed from two distinct promoters. No apparent sequence similarity was observed between the 5'-flanking regions of the two promoters. The expression of lck in human T-cell leukemia and carcinoma cell lines and in human peripheral blood T lymphocytes was examined by S1 nuclease and primer extension mapping and by Northern (RNA) blot analysis of total cellular RNA. The following results were obtained. (i) Two RNA start sites in the downstream promoter were used to generate type I transcripts. (ii) The major human type I start site has not been described for the mouse. (iii) At least five RNA start sites in the upstream promoter were used to generate type II transcripts. (iv) In T cells and in two colon carcinoma cell lines, type II transcripts were present in higher amounts than type I transcripts. (v) In T cells treated with phytohemagglutinin, tetradecanoylphorbol acetate, and cyclosporin A, the modulation of lck expression was associated primarily with changes in levels of type II transcripts. The above results suggest that the two human lck promoters are utilized differentially and may be regulated independently during certain physiological states.

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Apical beta 1 integrin in polarized MDCK cells mediates tubulocyst formation in response to type I collagen overlay

Journal of Cell Science ◽

10.1242/jcs.109.7.1875 ◽

1996 ◽

Vol 109 (7) ◽

pp. 1875-1889 ◽

Cited By ~ 3

Author(s):

A. Zuk ◽

K.S. Matlin

Keyword(s):

Plasma Membranes ◽

Type I Collagen ◽

Mdck Cells ◽

Basolateral Membrane ◽

Collagen Gel ◽

Type I ◽

Membrane Markers ◽

Polarized Epithelia ◽

Beta 1 Integrin ◽

Type I Collagen Gel

A number of epithelia form tubulocysts in vitro when overlaid with type I collagen gel. Because collagen receptors are generally believed to be expressed on the basolateral domain, the mechanism by which collagen elicits this morphogenetic response from the apical surface is unclear. To investigate the role of beta 1 integrins, the major receptor family for collagen, in this process, we overlaid polarized monolayers of MDCK II cells grown on permeable supports with type I collagen gel and correlated integrin polarity with the polarity of other apical and basolateral membrane markers during tubulocyst formation. Polarized monolayers of one clone of MDCK II cells, referred to as Heidelberg MDCK, initially respond to collagen overlay by stratifying; within 48 hours, lumena develop between the cell layers giving rise to tubulocysts. Tight junctions remain intact during tubulocyst formation because transepithelial electrical resistance does not significantly change. Major alterations are observed, however, in the expression and localization of apical and basolateral membrane markers. beta 1 integrins are necessary for tubulocyst morphogenesis because a function-blocking antibody administered to the apical pole of the cells completely inhibits the formation of these structures. To determine how apical-cell collagen interactions elicit tubulocyst formation, we examined whether beta 1 integrins are mobilized to apical plasma membranes in response to collagen overlay. We found that in the absence of collagen, polarized monolayers of Heidelberg MDCK cells endogenously express on apical plasma membranes a small pool of the beta 1 family, including alpha 2 beta 1 and alpha 3 beta 1. Collagen overlay does not mobilize additional beta 1 integrins to apical domains. If beta 1 integrins are not already apically expressed, as in the C6 MDCK cell line (Schoenenberger et al. (1994) J. Cell Biol. 107, 527–541), beta 1 integrins are not directed apically and tubulocysts do not develop in response to collagen. Thus, interaction of beta 1 integrin pre-existing on apical plasma membranes of polarized epithelia with type I collagen gel is the mechanism by which apical application of collagen elicits the formation of tubulocysts. Depolarized integrins on apical plasma membranes of polarized epithelia may be relevant to the pathogenesis of disease and injury.

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