Multiple functions of reversine on the biological characteristics of sheep fibroblasts

Abstract

Previous reports have demonstrated that Reversine can reverse differentiation of lineage-committed cells to mesenchymal stem cells and suppress tumors growth. However, the molecular mechanisms of antitumor activity and promoting cellular dedifferentiation for reversine have not yet been clearly elucidated. In the present study, it was demonstrated that reversine of 5 μM could induce multinucleated cells through cytokinesis failure rather than just arrested in G2 or M phase. Moreover, reversine reversed the differentiation of sheep fibroblasts into MSC-like style, and notably increased the expression of pluripotent marker genes Oct4 and MSCs-related surface antigens. The fibroblasts treated with reversine could transdifferentiate into all three germ layers cells in vitro. Most importantly, the induced β-like cells and hepatocytes had similar metabolic functions with normal cells in vivo. In addition, reversine promoted fibroblasts autophagy, ROS accumulation, mitochondrial dysfunction and cell apoptosis via the mitochondria mediated intrinsic pathway. The results of high-throughput RNA sequencing showed that most differentially expressed genes (DEGs) involved in Mismatch repair, Nucleotide excision repair and Base excision repair were significantly up-regulated in reversine treated fibroblasts, which means that high concentration of reversine will cause DNA damage and activate the DNA repair mechanism. In summary, reversine can increase the plasticity of sheep fibroblasts and suppress cell growth via the mitochondria mediated intrinsic pathway.

Reversine ameliorates hallmarks of cellular senescence in human skeletal myoblasts via reactivation of autophagy

“…Also, reversine decreased DNA damage, a hallmark of cellular senescence (López‐Otín et al, 2013), after only 4 days of treatment. On the contrary, SA‐β‐Gal expression, cell size and shape were restored 8–12 days after reversine treatment, indicating major reorganization of the actin cytoskeleton in agreement with previous work (Anastasia et al, 2006; Guo et al, 2021; Li et al, 2016). In agreement with previous reports where reversine was shown to inhibit cell proliferation (Huang et al, 2016; Park et al, 2019; Xia et al, 2021), cells did not proliferate during reversine treatment, likely due to inhibition of Aurora B kinase, but they started to proliferate after reversine was removed.…”

“…Several studies found that the small molecule 2,6-disubstituted purine, reversine, increased cellular plasticity as demonstrated by increased differentiation potential of progenitor cells toward the neuroectodermal lineage (Lee et al, 2009); dedifferentiation of C2C12 myoblasts to a progenitor-like state (Chen et al, 2004(Chen et al, , 2007; as well as dedifferentiation of sheep fibroblasts into multipotent progenitor cells, possibly via expression of the pluripotent factor, Oct4 (Guo et al, 2021). These progenitors could be induced to differentiate into adipocytes, osteoblasts, hepatocytes, and neurocytes in vitro (Guo et al, 2021). Other studies reported that reversine may have anticancer properties (D'Alise et al, 2008;Piccoli et al, 2020).…”

Association between reversine dose and increased plasticity of dedifferentiated fat (DFAT cells) into cardiac derived cells

“…Yu Guo et al showed that reversine can provoke cell cycle arrest, ROS accumulation, and mitochondrial dysfunction in DFAT cells triggering the apoptotic process. 29 It was also stated that reversine can induce apoptosis of DFAT cells through the mitochondrial-mediated intrinsic pathway, which has shown similar effects in all types of mesenchymal cells. 30 This indicates that toxic dose of reversine induces cell cycle termination and promotes cell apoptosis of DFAT differentiation.…”

Therapeutic function of a novel rat induced pluripotent stem cell line in a 6‑OHDA‑induced rat model of Parkinson's disease

“…Primary REFs were mechanically isolated and cultured from 16 embryonic day (E)16-18 rat embryos as previously described (29). Briefly, uteri were isolated from two E16-18 pregnant Sd rats following deep anesthesia via a subcutaneous injection of 3% sodium pentobarbital (50 mg/kg), as aforementioned.…”