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Updated: Jun 6, 2026
A Robust Discovery Platform for the Identification of Novel Mediators of Melanoma Metastasis
Published on: March 8, 2022
Variability in melanoma metalloproteinase expression profiling
Orsi Giricz1, Janelle L Lauer, Gregg B Fields
1Department of Chemistry and Biochemistry, Florida Atlantic University, Boca Raton, Florida 33431-0991, USA.
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.
Insights
Metalloproteinase gene expression, including matrix metalloproteinases (MMPs), is elevated in melanoma. Cultured cells show higher expression than tissue, highlighting the need for careful interpretation of cell culture experiments.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Metalloproteinase families, including ADAMs, ADAMTSs, and MMPs, are crucial in normal and pathological processes.
- These enzymes are implicated in extracellular matrix degradation and bioactive molecule processing.
Purpose of the Study:
- To analyze the gene expression of ADAM, MMP, and ADAMTS families in normal and metastatic melanoma.
- To compare metalloproteinase mRNA levels between tumor tissue and cultured cells.
Main Methods:
- Real-time PCR was used to quantify mRNA levels of selected ADAM, MMP, and ADAMTS genes.
- Analysis was performed on RNA from normal fibroblasts, metastatic melanoma cell lines, and their corresponding tissue samples.
Main Results:
- Increased gene expression of MMP1, MMP8, MMP15, and ADAM15 was observed in melanoma tissue and cell cultures.
- Metalloproteinase mRNA levels were generally higher in cultured cells compared to their tissue of origin.
- Passage-dependent expression patterns were noted for MMP8 and MMP9 in melanoma cell lines.
Conclusions:
- Specific ADAM and MMP members show altered expression in melanoma.
- Cell culture conditions can significantly influence metalloproteinase expression, necessitating cautious interpretation of results.
- Findings underscore the importance of considering in vitro artifacts when studying metalloproteinases in cancer.
