Sage Journals: Discover world-class research

Abstract

Human umbilical cord-derived mesenchymal stem cells (hUC-MSC) have been considered as promising candidates for cell-based regeneration medicine. However, the application was limited to its poor in vitro proliferation ability against the huge demand of cells. MicroRNA plays important roles in the regulation of cell proliferation, apoptosis, and differentiation. The objective of this study is to explore the roles of miRNAs in regulating the in vitro proliferation of hUC-MSC and unveil their possible mechanism. In this study, we found that miR-26b-3p was significantly upregulated during serial in vitro passage of hUC-MSC and was correlated with cellular senescence and cell cycle genes. The overexpression of miR-26b-3p greatly inhibited the proliferation of hUC-MSC in vitro, which is indicated by 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay, cell cycle, and cell growth curve analyses. miR-26b-3p suppression partly rescued this phenotype by maintaining its proliferation ability in vitro. For mechanism studies, we predicted and validated that miR-26b-3p suppresses estrogen receptor 1 (ESR1) expression by directly binding to the coding sequence (CDS) region of its message RNA (mRNA), thus subsequently changing the expression of its downstream effector Cyclin D1. In conclusion, we found that miR-26b-3p played an important role in the regulation of hUC-MSC proliferation in vitro by targeting the ESR-CCND1 pathway.

Get full access to this article

View all access options for this article.

References

Wang

, Qu

and Zhao

. (2012). Clinical applications of mesenchymal stem cells. J Hematol Oncol, 5:19.

Mundra

, Gerling

and Mahato

. (2013). Mesenchymal stem cell-based therapy. Mol Pharm, 10:77–89.

Caplan

and Correa

. The MSC: an injury drugstore. Cell Stem Cell, 9:11–15.

Yim

, Lee

, Bow

, Meij

, Leung

, Cheung

, Vavken

and Samartzis

. (2014). A systematic review of the safety and efficacy of mesenchymal stem cells for disc degeneration: insights and future directions for regenerative therapeutics. Stem Cells Dev, 23:2553–2567.

EI Omar

, Beroud

, Stoltz

, Menu

, Velot

and Decot

. (2014). Umbilical cord mesenchymal stem cells: the new gold standard for mesenchymal stem cell-based therapies?. Tissue Eng Part B Rev, 20:523–544.

Ikebe

and Suzuki

. (2014). Mesenchymal stem cells for regenerative therapy: optimization of cell preparation protocols. Biomed Res Int, 2014:951512.

Bartel

. (2009). MicroRNAs: target recognition and regulatory functions. Cell, 136:215–233.

Friedman

, Farh

, Burge

and Bartel

. (2009). Most mammalian mRNAs are conserved targets of microRNAs. Genome Res, 19:92–105.

Clark

, Kalomoiris

, Nolta

and Fierro

. (2014). Concise review: MicroRNA function in multipotent mesenchymal stromal cells. Stem Cells, 32:1074–1082.

10.

Trompeter

, Dreesen

, Hermann

, Iwaniuk

, Hafner

, Renwick

, Tuschl

and Wernet

. (2013). MicroRNAs miR-26a, miR-26b, and miR-29b accelerate osteogenic differentiation of unrestricted somatic stem cells from human cord blood. BMC Genomics, 14:111.

11.

Zhao

, Jiang

, Liu

, Xiang

, Zhou

and Chen

. (2015). MiR-124 promotes bone marrow mesenchymal stem cells differentiation into neurogenic cells for accelerating recovery in the spinal cord injury. Tissue Cell, 47:140–146.

12.

Yang

, Guo

, Zhang

, Chen

, Ying

and Dong

. (2011). MicroRNA-145 regulates chondrogenic differentiation of mesenchymal stem cells by targeting Sox9. PLoS One, 6:e21679.

13.

Hamam

, Ali

, Vishnubalaji

, Hamam

, Al-Nbaheen

, Chen

, Kassem

, Aldahmash

and Alajez

. (2014). microRNA-320/RUNX2 axis regulates adipocytic differentiation of human mesenchymal (skeletal) stem cells. Cell Death Dis, 5:e1499.

14.

Duan

, Sun

, Li

, Huang

, Xu

, Han

, Xing

and Yan

. (2014). miR-29a modulates neuronal differentiation through targeting REST in mesenchymal stem cells. PLoS One, 9:e97684.

15.

Zhang

, Xie

, Gordon

, LeBlanc

, Stein

, Lian

, van Wijnen

and Stein

. (2012). Control of mesenchymal lineage progression by microRNAs targeting skeletal gene regulators Trps1 and Runx2. J Biol Chem, 287:21926–21935.

16.

Zhang

, Wang

and Lü

. (2014). An integrated study of natural hydroxyapatite-induced osteogenic differentiation of mesenchymal stem cells using transcriptomics, proteomics and microRNA analyses. Biomed Mater, 9:045005.

17.

Hamam

, Ali

, Kassem

, Aldahmash

and Alajez

. (2015). microRNAs as regulators of adipogenic differentiation of mesenchymal stem cells. Stem Cells Dev, 24:417–425.

18.

Lin

, Zhang

, Huang

, et al. (2015). MiR-26b/KPNA2 axis inhibits epithelial ovarian carcinoma proliferation and metastasis through downregulating OCT4. Oncotarget, 6:23793–23806.

19.

, Sun

, Han

, Yu

, Hu

and Hu

. (2015). MiR-26b inhibits hepatocellular carcinoma cell proliferation, migration, and invasion by targeting EphA2. Int J Clin Exp Pathol, 8:4782–4790.

20.

Sun

, Yan

, Chen

, Yang

, Xu

, Mao

and Qiu

. (2015). MicroRNA-26b inhibits cell proliferation and cytokine secretion in human RASF cells via the Wnt/GSK-3β/β-catenin pathway. Diagn Pathol, 10:72.

21.

, Li

, Kong

, Luo

, Zhang

and Fang

. (2014). MiRNA-26b inhibits cellular proliferation by targeting CDK8 in breast cancer. Int J Clin Exp Med, 7:558–565.

22.

, Ji

, Song

, Shi

, Shen

, Chen

, Yang

, Zhao

and Guo

. (2015). Obesity-associated microRNA-26b regulates the proliferation of human preadipocytes via arrest of the G1/S transition. Mol Med Rep, 12:3648–3654.

23.

Das

, Sundell

and Koka

. (2013). Adult mesenchymal stem cells and their potency in the cell-based therapy. J Stem Cells, 8:1–16.

24.

Rehman

. (2010). Empowering self-renewal and differentiation: the role of mitochondria in stem cells. J Mol Med (Berl), 88:981–986.

25.

Wagers

. (2012). The stem cell niche in regenerative medicine. Cell Stem Cell, 10:362–369.

26.

Tsai

, Su

, Huang

, Yew

and Hung

. (2012). Oct4 and Nanog directly regulate Dnmt1 to maintain self-renewal and undifferentiated state in mesenchymal stem cells. Mol Cell, 47:169–182.

27.

Tome

, Lopez-Romero

, Albo

, Sepulveda

, Fernandez-Gutierrez

, Dopazo

, Bernad

and Gonzalez

. (2011). miR-335 orchestrates cell proliferation, migration and differentiation in human mesenchymal stem cells. Cell Death Differ, 18:985–995.

28.

Garofalo

, Di Leva

and Romano

. (2009). miR-221&222 regulate TRAIL resistance and enhance tumorigenicity through PTEN andTIMP3 downregulation. Cancer Cell, 16:498–509.

29.

Damdimopoulou

, Rodin

, Stenfelt

, Antonsson

, Tryggvason

and Hovatta

. (2015). Human embryonic stem cells. Best Pract Res Clin Obstet Gynaecol. S1521–S6934.

30.

Hou

, Niu

, Liu

, et al. (2015). Establishment and characterization of human germline stem cell line with unlimited proliferation potentials and no tumor formation. Sci Rep, 5:16922.

31.

Kamanga-Sollo

, White

, Hathaway

, Chung

, Johnson

and Dayton

. (2008). Roles of IGF-I and the estrogen, androgen and IGF-I receptors in estradiol-17beta- and trenbolone acetate-stimulated proliferation of cultured bovine satellite cells. Domest Anim Endocrinol, 35:88–97.

32.

Kamanga-Sollo

, White

, Weber

and Dayton

. (2013). Role of estrogen receptor-α (ESR1) and the type 1 insulin-like growth factor receptor (IGFR1) in estradiol-stimulated proliferation of cultured bovine satellite cells. Domest Anim Endocrinol, 44:36–45.

33.

Hong

, Zhang

, Sultana

, Yu

and Wei

. (2011). The effects of 17-β estradiol on enhancing proliferation of human bone marrow mesenchymal stromal cells in vitro. Stem Cells Dev, 20:925–931.

34.

Holzer

, Einhorn

and Majeska

. (2002). Estrogen regulation of growth and alkaline phosphatase expression by cultured human bone marrow stromal cells. J Orthop Res, 20:281–288.

35.

DiSilvio

, Jameson

, Gamie

, Giannoudis

and Tsiridis

. (2006). In vitro evaluation of the direct effect of estradiol on human osteoblasts (HOB) and human mesenchymal stem cells (h-MSCs). Injury 37, (Suppl. 3)::S33–S42.

36.

Chen

, Hu

and Wang

. (2013). Estrogen modulates osteogenic activity and estrogen receptor mRNA in mesenchymal stem cells of women. Climacteric, 16:154–160.

37.

Glass

. (1994). Differential recognition of target genes by nuclear receptor monomers, dimers, and heterodimers. Endocr Rev, 15:391–407.

38.

Beato

, Herrlich

and Schütz

. (1995). Steroid hormone receptors: many actors in search of a plot. Cell, 83:851–857.

39.

Rickard

, Subramaniam

and Spelsberg

. (1999). Molecular and cellular mechanisms of estrogen action on the skeleton. J Cell Biochem Suppl, 32–33:123–132.

40.

Gao

and Dahlman-Wright

. (2010). The gene regulatory networks controlled by estrogens. Mol Cell Endocrinol, 334:83–90.

41.

Carroll

, Liu

, Brodsky

, et al. (2005). Chromosome-wide mapping of estrogen receptor binding reveals long-range regulation requiring the forkhead protein FoxA1. Cell, 122:33–43.

42.

Lucas

, Lazari

and Porto

. (2014). Differential role of the estrogen receptors ESR1 and ESR2 on the regulation of proteins involved with proliferation and differentiation of Sertoli cells from 15-day-old rats. Mol Cell Endocrinol, 382:84–96.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

0.06 MB

0.02 MB

miR-26b-3p Regulates Human Umbilical Cord-Derived Mesenchymal Stem Cell Proliferation by Targeting Estrogen Receptor

Abstract

Get full access to this article

References

Supplementary Material