Epithelial to mesenchymal transition in mammary gland tissue fibrosis and insights into drug therapeutics

Mudasir Ahmad Syed; Basharat Bhat; Abiza Wali; Afnan Saleem; Lateef Ahmad Dar; Mudasir Bashir Gugjoo; Shakil Bhat; Sahar Saleem Bhat

doi:10.7717/peerj.15207

Epithelial to mesenchymal transition in mammary gland tissue fibrosis and insights into drug therapeutics

PeerJ. 2023 May 9:11:e15207. doi: 10.7717/peerj.15207. eCollection 2023.

Authors

Mudasir Ahmad Syed¹, Basharat Bhat¹, Abiza Wali², Afnan Saleem¹, Lateef Ahmad Dar¹, Mudasir Bashir Gugjoo³, Shakil Bhat¹, Sahar Saleem Bhat¹

Affiliations

¹ Division of Animal Biotechnology, Faculty of Veterinary Sciences, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, India, Srinagar, India.
² Department of Clinical Biochemistry, University of Kashmir, Srinagar, Jammu and Kashmir, India.
³ Division of Veterinary Surgery, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Faculty of Veterinary Sciences and Animal Husbandry, Shuhama, SKUAST-K, India, Srinagar, Jammu and Kashmir, India.

Abstract

Background: The epithelial-mesenchymal transition (EMT) is a multi-step morphogenetic process in which epithelial cells lose their epithelial properties and gain mesenchymal characteristics. The process of EMT has been shown to mediate mammary gland fibrosis. Understanding how mesenchymal cells emerge from an epithelial default state will aid in unravelling the mechanisms that control fibrosis and, ultimately, in identifying therapeutic targets to alleviate fibrosis.

Methods: The effects of EGF and high glucose (HG) on EMT in mammary epithelial cells, MCF10A and GMECs, as well as their pathogenic role, were studied. In-silico analysis was used to find interacting partners and protein-chemical/drug molecule interactions.

Results: On treatment with EGF and/or HG, qPCR analysis showed a significant increase in the gene expression of EMT markers and downstream signalling genes. The expression of these genes was reduced on treatment with EGF+HG combination in both cell lines. The protein expression of COL1A1 increased as compared to the control in cells treated with EGF or HG alone, but when the cells were treated with EGF and HG together, the protein expression of COL1A1 decreased. ROS levels and cell death increased in cells treated with EGF and HG alone, whereas cells treated with EGF and HG together showed a decrease in ROS production and apoptosis. In-silico analysis of protein-protein interactions suggest the possible role of MAPK1, actin alpha 2 (ACTA2), COL1A1, and NFκB1 in regulating TGFβ1, ubiquitin C (UBC), specificity protein 1 (SP1) and E1A binding protein P300 (EP300). Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment suggests advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signalling pathway, relaxin signalling pathway and extra cellular matrix (ECM) receptor interactions underlying fibrosis mechanism.

Conclusion: This study demonstrates that EGF and HG induce EMT in mammary epithelial cells and may also have a role in fibrosis.

Keywords: Drug targeting; Epithelial growth factor; Epithelium-mesenchymal transition; Fibrosis; Goat mammary epithelial cells; MCF10A.

MeSH terms

Epidermal Growth Factor / pharmacology
Epithelial-Mesenchymal Transition* / genetics
Fibrosis
Humans
Mammary Glands, Human*
Reactive Oxygen Species / pharmacology

Substances

Epidermal Growth Factor
Reactive Oxygen Species

Grants and funding

The authors received no funding for this work.