The Effect of Bta-miR-1296 on the Proliferation and Extracellular Matrix Synthesis of Bovine Mammary Fibroblasts

Overview

Journal In Vitro Cell Dev Biol Anim

Publisher Springer

Specialties Biology
Cell Biology

Date 2024 Feb 27

PMID 38409638

Authors

Yuejie Yang

Tao Yuan

Rui Wu

Zijian Geng

Shuai Lian

Jianfa Wang

Affiliations

Soon will be listed here.

Abstract

Mammary fibrosis in dairy cows is a chronic condition caused by mastitis, and can lead to serious culling of dairy cows resulting in huge economic losses in the dairy industry. MicroRNAs (miRNAs) exert an important role in regulating mammary gland health in dairy cows. This study investigated whether exosomal miRNAs in mammary epithelial cells can regulate the proliferation of bovine mammary fibroblasts (BMFBs) in mastitis. Liposome transfection technology was used to construct a cellular model of the overexpression and inhibition of miRNAs. The STarMir software, dual luciferase reporter gene test, real-time quantitative PCR (qRT-PCR), a Cell Counting Kit-8 (CCK-8), and a Western Blot and plate clone formation test were used to investigate the mechanism by which bta-miR-1296 regulates the proliferation of BMFBs. Target gene prediction results revealed that glutamate-ammonia ligase was a direct target gene by which bta-miR-1296 regulates cell proliferation. It was found that bta-miR-1296 significantly inhibited the proliferation of BMFBs. After BMFBs were transfected with a bta-miR-1296 mimic, mRNA expression in the extracellular matrix (ECM), α-smooth muscle actin (α-SMA), collagen type I alpha 1 chain (COL1α1) and collagen type III alpha 1 chain (COL3α1), and various cell growth factors (basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor-BB (PDGF-BB), and transforming growth factor-β1 (TGF-β1)) were down-regulated, and the expressions of α-SMA, COL1α1, COL3α1, phospho-extracellular regulated protein kinases, phospho-protein kinase B, TGF-β1, and phospho-Smad family member3 proteins were inhibited. In conclusion, bta-miR-1296 can inhibit the proliferation of BMFBs and the synthesis of ECM in BMFBs, thus affecting the occurrence and development of mammary fibrosis in dairy cows and laying the foundation for further studies to clarify the regulatory mechanism of mammary fibrosis.

References

Barratt S, Blythe T, Ourradi K, Jarrett C, Welsh G, Bates D . Effects of hypoxia and hyperoxia on the differential expression of VEGF-A isoforms and receptors in Idiopathic Pulmonary Fibrosis (IPF). Respir Res. 2018; 19(1):9. PMC: 5769544. DOI: 10.1186/s12931-017-0711-x. View

Bi Y, Ding Y, Wu J, Miao Z, Wang J, Wang F . Staphylococcus aureus induces mammary gland fibrosis through activating the TLR/NF-κB and TLR/AP-1 signaling pathways in mice. Microb Pathog. 2020; 148:104427. DOI: 10.1016/j.micpath.2020.104427. View

Buckley C . Why does chronic inflammation persist: An unexpected role for fibroblasts. Immunol Lett. 2011; 138(1):12-4. PMC: 3110265. DOI: 10.1016/j.imlet.2011.02.010. View

Chen G, He M, Yin Y, Yan T, Cheng W, Huang Z . miR-1296-5p decreases ERBB2 expression to inhibit the cell proliferation in ERBB2-positive breast cancer. Cancer Cell Int. 2017; 17:95. PMC: 5655974. DOI: 10.1186/s12935-017-0466-y. View

Chen Q, Yang W, Wang X, Li X, Qi S, Zhang Y . TGF-β1 Induces EMT in Bovine Mammary Epithelial Cells Through the TGFβ1/Smad Signaling Pathway. Cell Physiol Biochem. 2017; 43(1):82-93. DOI: 10.1159/000480321. View

Choi J, Lim K, Park E, Kim J, Choi Y, Baek K . Glutamate-ammonia ligase and reduction of G0 population in PANC-1 cells. J Cell Biochem. 2012; 114(2):303-13. DOI: 10.1002/jcb.24370. View

Deng H, Xie C, Ye Y, Du Z . MicroRNA-1296 expression is associated with prognosis and inhibits cell proliferation and invasion by Wnt signaling in non-small cell lung cancer. Oncol Lett. 2020; 19(1):623-630. PMC: 6924095. DOI: 10.3892/ol.2019.11154. View

Feng X, Derynck R . Specificity and versatility in tgf-beta signaling through Smads. Annu Rev Cell Dev Biol. 2005; 21:659-93. DOI: 10.1146/annurev.cellbio.21.022404.142018. View

Gao Y, Wang Y, Li Y, Xia X, Zhao S, Che Y . TGF-β1 promotes bovine mammary fibroblast proliferation through the ERK 1/2 signalling pathway. Cell Biol Int. 2016; 40(7):750-60. DOI: 10.1002/cbin.10609. View

10.

Gjorevski N, Nelson C . Integrated morphodynamic signalling of the mammary gland. Nat Rev Mol Cell Biol. 2011; 12(9):581-93. DOI: 10.1038/nrm3168. View

11.

He G, Ma M, Yang W, Wang H, Zhang Y, Gao M . SDF-1 in Mammary Fibroblasts of Bovine with Mastitis Induces EMT and Inflammatory Response of Epithelial Cells. Int J Biol Sci. 2017; 13(5):604-614. PMC: 5441177. DOI: 10.7150/ijbs.19591. View

12.

Henderson N, Rieder F, Wynn T . Fibrosis: from mechanisms to medicines. Nature. 2020; 587(7835):555-566. PMC: 8034822. DOI: 10.1038/s41586-020-2938-9. View

13.

Iwano M, Plieth D, Danoff T, Xue C, Okada H, Neilson E . Evidence that fibroblasts derive from epithelium during tissue fibrosis. J Clin Invest. 2002; 110(3):341-50. PMC: 151091. DOI: 10.1172/JCI15518. View

14.

Maseki S, Ijichi K, Tanaka H, Fujii M, Hasegawa Y, Ogawa T . Acquisition of EMT phenotype in the gefitinib-resistant cells of a head and neck squamous cell carcinoma cell line through Akt/GSK-3β/snail signalling pathway. Br J Cancer. 2012; 106(6):1196-204. PMC: 3304404. DOI: 10.1038/bjc.2012.24. View

15.

Massague J, Gomis R . The logic of TGFbeta signaling. FEBS Lett. 2006; 580(12):2811-20. DOI: 10.1016/j.febslet.2006.04.033. View

16.

Schlessinger J . Common and distinct elements in cellular signaling via EGF and FGF receptors. Science. 2004; 306(5701):1506-7. DOI: 10.1126/science.1105396. View

17.

Scotton C, Chambers R . Molecular targets in pulmonary fibrosis: the myofibroblast in focus. Chest. 2007; 132(4):1311-21. DOI: 10.1378/chest.06-2568. View

18.

Seki E, De Minicis S, Osterreicher C, Kluwe J, Osawa Y, Brenner D . TLR4 enhances TGF-beta signaling and hepatic fibrosis. Nat Med. 2007; 13(11):1324-32. DOI: 10.1038/nm1663. View

19.

Shan X, Wen W, Zhu D, Yan T, Cheng W, Huang Z . miR 1296-5p Inhibits the Migration and Invasion of Gastric Cancer Cells by Repressing ERBB2 Expression. PLoS One. 2017; 12(1):e0170298. PMC: 5242522. DOI: 10.1371/journal.pone.0170298. View

20.

Tao Y, Ma C, Fan Q, Wang Y, Han T, Sun C . MicroRNA-1296 Facilitates Proliferation, Migration And Invasion Of Colorectal Cancer Cells By Targeting SFPQ. J Cancer. 2018; 9(13):2317-2326. PMC: 6036719. DOI: 10.7150/jca.25427. View