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

Mol Biol Rep. 2022 May;49(5):3665-3674. doi: 10.1007/s11033-022-07206-6. Epub 2022 Feb 2.

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.

Keywords: Metallothionein; Mouth neoplasms; Squamous cell carcinoma; Tumour.

MeSH terms

  • Carcinoma, Squamous Cell* / pathology
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Cell Proliferation / genetics
  • Down-Regulation / genetics
  • Epidermal Growth Factor / genetics
  • Humans
  • Ki-67 Antigen / metabolism
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase 9* / genetics
  • Metallothionein* / genetics
  • Transforming Growth Factor alpha / genetics
  • Tumor Necrosis Factor-alpha / genetics

Substances

  • Ki-67 Antigen
  • MT2A protein, human
  • Transforming Growth Factor alpha
  • Tumor Necrosis Factor-alpha
  • Epidermal Growth Factor
  • Metallothionein
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9