Research Paper Volume 16, Issue 5 pp 4904—4919
MicroRNA-18a prevents senescence of mesenchymal stem cells by targeting CTDSPL
- 1 State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China
- 2 Department of Clinical Science and Research, Zhongda Hospital, School of Medicine, Southeast University, Nanjing 210009, China
Received: August 29, 2023 Accepted: December 26, 2023 Published: March 7, 2024
https://doi.org/10.18632/aging.205642How to Cite
Copyright: © 2024 Sun et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Stem cell therapy requires massive-scale homogeneous stem cells under strict qualification control. However, Prolonged ex vivo expansion impairs the biological functions and results in senescence of mesenchymal stem cells (MSCs). We investigated the function of CTDSPL in the premature senescence process of MSCs and clarified that miR-18a-5p played a prominent role in preventing senescence of long-term cultured MSCs and promoting the self-renewal ability of MSCs. Over-expression of CTDSPL resulted in an enlarged morphology, up-regulation of p16 and accumulation of SA-β-gal of MSCs. The reduced phosphorylated RB suggested cell cycle arrest of MSCs. All these results implied that CTDSPL induced premature senescence of MSCs. We further demonstrated that miR-18a-5p was a putative regulator of CTDSPL by luciferase reporter assay. Inhibition of miR-18a-5p promoted the expression of CTDSPL and induced premature senescence of MSCs. Continuous overexpression of miR-18a-5p improved self-renewal of MSCs by reducing ROS level, increased expression of Oct4 and Nanog, and promoted growth rate and differentiation capability. We reported for the first time that the dynamic interaction of miR-18a-5p and CTDSPL is crucial for stem cell senescence.
Abbreviations
ALP: Alkaline phosphatase; BSA: Bovine serum albumin; CTDSPL: Carboxy-terminal domain RNA polymerase II polypeptide A small phosphatase-like; ECL: Enhanced chemiluminescence substrate; ECL: Enhanced chemiluminescence substrate; FBS: Fetal bovine serum; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; GFP: Green fluorescent protein; MSCs: Mesenchymal stem cells; PDT: Population double time; PFA: Paraformaldehyde; RB: Retinoblastoma gene; ROS: Reactive oxygen species; SA-β-gal: Senescence-associated β-galactosidase; UCMSCs: Mesenchymal stem cells derived from human umbilical cord; UTR: Untranslated region.