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Downregulation of metallothionein 2A reduces migration, invasion and proliferation activities in human squamous cell carcinoma cells

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Abstract

Background

The invasive behaviour of squamous cell carcinoma (SCC), a common malignant tumour of the mouth, is a process mediated by cell proliferation, extracellular matrix proteolysis and other factors. Studies have shown a potential relationship between growth factors, metallothionein 2A (MT2A) and matrix metalloproteinase (MMP) activation in malignant tumours. The aim of this study was to downregulate MT2A in cells (Cal27) derived from human squamous cell carcinoma.

Methods

Cal27 cells with reduced MT2A were subjected to proliferation, migration and invasion assays. Immunofluorescence and western blot confirmed MT2A depletion by siRNA. Growth curve assays assessed cell proliferation. Indirect immunofluorescence analysed the expression of MT2A, MMP-2, MMP-9, epidermal growth factor (EGF), transforming growth factor alpha (TGF-α), tumour necrosis factor alpha (TNF-α) and Ki67. Zymography evaluated the effects of MT2A silencing on MMP-2 and -9 expression. Migration and invasion activities were evaluated using migration and invasion assays.

Results

CAL27 cells displayed MT2A, MMP-2, MMP-9, EGF, TGF-α, TNF-α and Ki67. MT2A depletion decreased MMP-9, EGF, TGF-α and Ki67 protein levels, while increasing TNF-α.

Conclusions

MT2A downregulation reduced cell proliferation, migration and invasion activities. Therefore, MT2A has an important role in cell proliferation, migration and invasion in human oral SCC cells.

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Data Availability

The datasets generated or analysed during the current study are available from the corresponding author on reasonable request. Supplementary information is available for this paper (see Supplementary Information file).

Abbreviations

EGF:

Epidermal growth factor

ECM:

Extracellular matrix

FBS:

Foetal bovine serum

MMP:

Matrix metalloproteinase

MT:

Metallothionein

MT2A:

Metallothionein 2A

SCC:

Squamous cell carcinoma

TGF-α:

Transforming growth factor alpha

TNF-α:

Tumour necrosis factor alpha

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Acknowledgements

Not applicable.

Funding

This research was funded by FAPESPA (Foundation for Research Support of the State of Pará) linked to the State Secretariat for Development, Science and Technology, grant number 006/2016.

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Author notes

  1. João de Jesus Viana Pinheiro and Sérgio de Melo Alves Junior have contributed equally to this work.

Authors and Affiliations

  1. Department of Oral Pathology, School of Dentistry, Universidade Federal do Para, Avenida Augusto Corrêa, 01, Belém, PA, 66075-110, Brazil

    Aline Marques Dias, Raíssa Pinheiro de Mendonça, Maria Sueli da Silva Kataoka, João de Jesus Viana Pinheiro & Sérgio de Melo Alves Junior

  2. Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of Sao Paulo, Av. Prof. Lineu Prestes 1524, Sao Paulo, 05508-000, Brazil

    Ruy G. Jaeger

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Contributions

J.J.V.P. and S.M.A.J performed the conceptualization, formal analysis and project administration. M.S.S.K., J.J.V.P. and S.M.A.J. defined the methodology and provided resources. M.S.S.K. and J.J.V.P. validated the project. A.M.D., R.P.M, and R. G. J. performed the investigation and wrote the main manuscript text. A.M.D., J.J.V.P. and S.M.A.J. were responsible for data curation and funding acquisition. All authors reviewed and edited the manuscript.

Corresponding author

Correspondence to João de Jesus Viana Pinheiro.

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This research was conducted in full accordance with ethical principles and was approved by the Ethics Committee on Research in Human Beings of the Institute of Health Sciences of the Federal University of Pará (Protocol Number 1.593.119).

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Dias, A.M., de Mendonça, R.P., da Silva Kataoka, M.S. et al. Downregulation of metallothionein 2A reduces migration, invasion and proliferation activities in human squamous cell carcinoma cells. Mol Biol Rep 49, 3665–3674 (2022). https://doi.org/10.1007/s11033-022-07206-6

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  • DOI: https://doi.org/10.1007/s11033-022-07206-6

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