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MGSA/GRO transcription is differentially regulated in normal retinal pigment epithelial and melanoma cells.

Publication ,  Journal Article
Shattuck, RL; Wood, LD; Jaffe, GJ; Richmond, A
Published in: Mol Cell Biol
January 1994

We have characterized constitutive and cytokine-regulated MGSA/GRO alpha, -beta, and -gamma gene expression in normal retinal pigment epithelial (RPE) cells and a malignant melanoma cell line (Hs294T) to discern the mechanism for MGSA/GRO constitutive expression in melanoma. In RPE cells, constitutive MGSA/GRO alpha, -beta, and -gamma mRNAs are not detected by Northern (RNA) blot analysis although nuclear runoff experiments show that all three genes are transcribed. In Hs294T cells, constitutive MGSA/GRO alpha expression is detectable by Northern blot analysis, and the level of basal MGSA/GRO alpha transcription is 8- to 30-fold higher than in RPE cells. In contrast, in Hs294T cells, basal MGSA/GRO beta and -gamma transcription is only twofold higher than in RPE cells and no beta or gamma mRNA is detected by Northern blot. These data suggest that the constitutive MGSA/GRO alpha mRNA in Hs294T cells is due to increased basal MGSA/GRO alpha gene transcription. The cytokines interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF alpha) significantly increase the mRNA levels for all three MGSA/GRO isoforms in Hs294T and RPE cells, and both transcriptional and posttranscriptional mechanisms are operational. Nuclear runoff assays indicate that in RPE cells, a 1-h IL-1 treatment induces a 10- to 20-fold increase in transcription of MGSA/GRO alpha, -beta and -gamma but only a 2-fold increase in Hs294T cells. Similarly, chloramphenicol acetyltransferase (CAT) reporter gene analysis using the MGSA/GRO alpha, -beta, and -gamma promoter regions demonstrates that IL-1 treatment induces an 8- to 14-fold increase in CAT activity in RPE cells but only a 2-fold increase in Hs294T cells. The effect of deletion or mutation of the MGSA/GRO alpha NF-kappa B element, combined with data from gel mobility shift analyses, indicates that the NF-kappa B p50/p65 heterodimer in RPE cells plays an important role in IL-1- and TNF alpha-enhanced gene transcription. In Hs294T cells, gel shift analyses indicate that IL-1 and TNF alpha induce NF-kappa B complex formation; however, transactivation does not occur, suggesting that subtle differences in the NF-kappa B complexes may result in the inability of the cytokines IL-1 and TNF alpha to activate transcription of the MGSA/GRO genes. IL-1 and TNF alpha posttranscriptionally regulate MGSA/GRO mRNA levels in both cell types. In Hs294T cells, IL-1 increases the half-life of MGSA/GRO alpha from 15 min to 6 h (a 24-fold increase in half-life). These data indicate that IL-1 and TNF alpha transcriptionally and posttranscriptionally regulate MGSA/GRO alpha, -beta, and -gamma mRNA levels in RPE cells, while in Hs294T cells, the major effect of IL-1 and TNF alpha is on mRNA stability.

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Published In

Mol Cell Biol

DOI

ISSN

0270-7306

Publication Date

January 1994

Volume

14

Issue

1

Start / End Page

791 / 802

Location

United States

Related Subject Headings

  • Tumor Necrosis Factor-alpha
  • Transfection
  • Transcription, Genetic
  • RNA, Messenger
  • Pigment Epithelium of Eye
  • Oligonucleotide Probes
  • Neoplasm Proteins
  • NF-kappa B
  • Molecular Sequence Data
  • Melanoma
 

Citation

APA
Chicago
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MLA
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Shattuck, R. L., Wood, L. D., Jaffe, G. J., & Richmond, A. (1994). MGSA/GRO transcription is differentially regulated in normal retinal pigment epithelial and melanoma cells. Mol Cell Biol, 14(1), 791–802. https://doi.org/10.1128/mcb.14.1.791-802.1994
Shattuck, R. L., L. D. Wood, G. J. Jaffe, and A. Richmond. “MGSA/GRO transcription is differentially regulated in normal retinal pigment epithelial and melanoma cells.Mol Cell Biol 14, no. 1 (January 1994): 791–802. https://doi.org/10.1128/mcb.14.1.791-802.1994.
Shattuck RL, Wood LD, Jaffe GJ, Richmond A. MGSA/GRO transcription is differentially regulated in normal retinal pigment epithelial and melanoma cells. Mol Cell Biol. 1994 Jan;14(1):791–802.
Shattuck, R. L., et al. “MGSA/GRO transcription is differentially regulated in normal retinal pigment epithelial and melanoma cells.Mol Cell Biol, vol. 14, no. 1, Jan. 1994, pp. 791–802. Pubmed, doi:10.1128/mcb.14.1.791-802.1994.
Shattuck RL, Wood LD, Jaffe GJ, Richmond A. MGSA/GRO transcription is differentially regulated in normal retinal pigment epithelial and melanoma cells. Mol Cell Biol. 1994 Jan;14(1):791–802.

Published In

Mol Cell Biol

DOI

ISSN

0270-7306

Publication Date

January 1994

Volume

14

Issue

1

Start / End Page

791 / 802

Location

United States

Related Subject Headings

  • Tumor Necrosis Factor-alpha
  • Transfection
  • Transcription, Genetic
  • RNA, Messenger
  • Pigment Epithelium of Eye
  • Oligonucleotide Probes
  • Neoplasm Proteins
  • NF-kappa B
  • Molecular Sequence Data
  • Melanoma