Epigenetic modifications of histone H3 during the transdifferentiation of Thy-1(+) Lin(‑) bone marrow cells into hepatocytes

Liao,Xinxin; Liao,Yixin; Zou,Yantai; Li,Guanhong; Liao,Caixian

doi:10.3892/mmr.2015.4384

Epigenetic modifications of histone H3 during the transdifferentiation of Thy-1(+) Lin(‑) bone marrow cells into hepatocytes

Authors:
- Xinxin Liao
- Yixin Liao
- Yantai Zou
- Guanhong Li
- Caixian Liao
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Affiliations: Department of Anesthesiology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Obstetrics and Gynecology, Center for Reproductive Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Hepatobiliary Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China
Published online on: September 29, 2015 https://doi.org/10.3892/mmr.2015.4384
Pages: 7561-7567

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Abstract

The epigenetic modifications during the transdifferentiation of adult stem cells remain to be fully elucidated. In the present study, the histone H3 modifications during the transdifferentiation of rat Thy‑1(+) Lin(‑) bone marrow cells into hepatocytes in vitro were examined, which involved performing hepatocyte growth factor-mediated transdifferentiation of bone marrow Thy-1(+) Lin(‑) cells into hepatic lineage cells. Subsequently, the hepatocyte-specific markers, cytokeratin‑18 (CK‑18), albumin (ALB) and α‑fetoprotein (AFP) were examined by immunofluorescence staining or reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Changes in the key pluripotency factor, octamer‑binding transcription factor 4 (OCT4) and histone modifications, including the dimethylation and acetylation of H3 at lysine 9 (H3K9me2 and H3K9ac), lysine 14 (H3K14me2 and H3K14ac) and lysine 27 (H3K27me2 and H3K27ac), were also investigated by RT-qPCR, immunofluorescence staining or western blot analysis The mRNA expression levels of AFP and ALB were detected in the bone marrow stem cell‑derived hepatic lineage cells on days 7 and 14 following induction, and CK‑18 was detected on day 14 following induction. During the transdifferentiation of the bone marrow Thy‑1(+) Lin(‑) cells into hepatocytes, the mRNA expression of OCT4 was significantly reduced, and the levels of H3K9me2, H3K9ac, H3K14me2, H3K14ac, H3K27me2 and H3K27ac were increased significantly, compared with the levels at baseline (P<0.05). Therefore, the results of the present study demonstrated that histone H3 modifications at lysine 9, 14 and 27 are involved in the regulation of transcription during the transdifferentiation of bone marrow stem cells to hepatic lineage cells.

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1	Filip S, English D and Mokrý J: Issues in stem cell plasticity. J Cell Mol Med. 8:572–577. 2004. View Article : Google Scholar : PubMed/NCBI
2	Tang Y, He H, Cheng N, Song Y, Ding W, Zhang Y, Zhang W, Zhang J, Peng H and Jiang H: PDGF, NT-3 and IGF-2 in combination induced transdifferentiation of muscle-derived stem cells into Schwann cell-like cells. PLoS One. 9:e734022014. View Article : Google Scholar : PubMed/NCBI
3	Snykers S, De Kock J, Rogiers V and Vanhaecke T: In vitro differentiation of embryonic and adult stem cells into hepatocytes: State of the art. Stem Cells. 27:577–605. 2009. View Article : Google Scholar :
4	Tang H, Liao CX, Zhou J, Jin HS, Tan YF, Su J, Zhang CX and Zhang SH: Differentiation of transplanted mouse c-Kit+lin-bone marrow cells into hepatocytes in vitro. Journal of Southern Medical University. 26:567–569. 2006.In Chinese.
5	Liao X, AnCheng JY, Zhou QJ and Liao C: Therapeutic effect of autologous bone marrow-derived liver stem cells transplantation in hepatitis B virus-induced liver cirrhosis. Hepatogastroenterology. 60:406–409. 2013.
6	Vaillant I and Paszkowski J: Role of histone and DNA methylation in gene regulation. Curr Opin Plant Biol. 10:528–533. 2007. View Article : Google Scholar : PubMed/NCBI
7	Allis CD, Berger SL, Cote J, Dent S, Jenuwien T, Kouzarides T, Pillus L, Reinberg D, Shi Y, Shiekhattar R, et al: New nomenclature for chromatin-modifying enzymes. Cell. 131:633–636. 2007. View Article : Google Scholar : PubMed/NCBI
8	Miao F and Natarajan R: Mapping global histone methylation patterns in the coding regions of human genes. Mol Cell Biol. 25:4650–4661. 2005. View Article : Google Scholar : PubMed/NCBI
9	Kouzarides T: Chromatin modifications and their function. Cell. 128:693–705. 2007. View Article : Google Scholar : PubMed/NCBI
10	Bannister AJ, Zegerman P, Partridge JF, Miska EA, Thomas JO, Allshire RC and Kouzarides T: Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain. Nature. 410:120–124. 2001. View Article : Google Scholar : PubMed/NCBI
11	Schones DE and Zhao K: Genome-wide approaches to studying chromatin modifications. Nat Rev Genet. 9:179–191. 2008. View Article : Google Scholar : PubMed/NCBI
12	Kim H, Jang MJ, Kang MJ and Han YM: Epigenetic signatures and temporal expression of lineage-specific genes in hESCs during differentiation to hepatocytes in vitro. Hum Mol Genet. 20:401–412. 2011. View Article : Google Scholar
13	Taverna SD, Li H, Ruthenburg AJ, Allis CD and Patel DJ: How chromatin-binding modules interpret histone modifications: Lessons from professional pocket pickers. Nat Struct Mol Biol. 14:1025–1040. 2007. View Article : Google Scholar : PubMed/NCBI
14	Chen Y, Pan RL, Zhang XL, Shao JZ, Xiang LX, Dong XJ and Zhang GR: Induction of hepatic differentiation of mouse bone marrow stromal stem cells by the histone deacetylase inhibitor VPA. J Cell Mol Med. 13:2582–2592. 2009. View Article : Google Scholar
15	Guittin P and Decelle T: Future improvements and implementation of animal care practices within the animal testing regulatory environment. ILAR J. 43(Suppl): S80–S84. 2002.PubMed/NCBI
16	Zhang CX, Liao SH, Zhang J, Sun J and Zhou: Separation of bone marrow-derived liver stem cell subset Thy-1.2 + Lin-cells using immunomagnetic beads. Shandong University Transaction (Medical Sciences). 46:108–110. 2008.
17	Oh SH, Miyazaki M, Kouchi H, Inoue Y, Sakaguchi M, Tsuji T, Shima N, Higashio K and Namba M: Hepatocyte growth factor induces differentiation of adult rat bone marrow cells into a hepatocyte lineage in vitro. Biochem Biophys Res Commun. 279:500–504. 2000. View Article : Google Scholar : PubMed/NCBI
18	Elyaman W, Yardin C and Hugon J: Involvement of glycogen synthase kinase-3beta and tau phosphorylation in neuronal Golgi disassembly. J Neurochem. 81:870–880. 2002. View Article : Google Scholar : PubMed/NCBI
19	Kang XQ, Zang WJ, Bao LJ, Li DL, Song TS, Xu XL and Yu XJ: Fibroblast growth factor-4 and hepatocyte growth factor induce differentiation of human umbilical cord blood-derived mesen-chymal stem cells into hepatocytes. World J Gastroenterol. 11:7461–7465. 2005. View Article : Google Scholar
20	Lee KD, Kuo TK, Whang-Peng J, Chung YF, Lin CT, Chou SH, Chen JR, Chen YP and Lee OK: In vitro hepatic differentiation of human mesenchymal stem cells. Hepatology. 40:1275–1284. 2004. View Article : Google Scholar : PubMed/NCBI
21	Kang XQ, Zang WJ, Song TS, Xu XL, Yu XJ, Li DL, Meng KW, Wu SL and Zhao ZY: Rat bone marrow mesenchymal stem cells differentiate into hepatocytes in vitro. World J Gastroenterol. 11:3479–84. 2005. View Article : Google Scholar : PubMed/NCBI
22	Ong SY, Dai H and Leong KW: Hepatic differentiation potential of commercially available human mesenchymal stem cells. Tissue Eng. 12:3477–3485. 2006. View Article : Google Scholar
23	Leu YW, Huang TH and Hsiao SH: Epigenetic reprogramming of mesenchymal stem cells. Adv Exp Med Biol. 754:195–211. 2013. View Article : Google Scholar
24	Yamanaka Y and Ralston A: Early embryonic cell fate decisions in the mouse. Adv Exp Med Biol. 695:1–13. 2010. View Article : Google Scholar
25	Chui HC and Damasio AR: Progressive dialysis encephalopathy (ʻdialysis dementia'). J Neurol. 222:145–157. 1980. View Article : Google Scholar : PubMed/NCBI
26	Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP, Guenther MG, Kumar RM, Murray HL, Jenner RG, et al: Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 122:947–956. 2005. View Article : Google Scholar : PubMed/NCBI
27	Nichols J, Zevnik B, Anastassiadis K, Niwa H, Klewe-Nebenius D, Chambers I, Schöler H and Smith A: Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell. 95:379–391. 1998. View Article : Google Scholar : PubMed/NCBI
28	Takahashi K and Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 126:663–676. 2006. View Article : Google Scholar : PubMed/NCBI
29	Athanasiadou R, de Sousa D, Myant K, Merusi C, Stancheva I and Bird A: Targeting of de novo DNA methylation throughout the Oct-4 gene regulatory region in differentiating embryonic stem cells. PLoS One. 5:e99372010. View Article : Google Scholar : PubMed/NCBI
30	Khanjani S, Khanmohammadi M, Zarnani AH, Talebi S, Edalatkhah H, Eghtesad S, Nikokar I and Kazemnejad S: Efficient generation of functional hepatocyte-like cells from menstrual blood-derived stem cells. J Tissue Eng Regen Med. 2013. View Article : Google Scholar : PubMed/NCBI
31	Turner BM: Reading signals on the nucleosome with a new nomenclature for modified histones. Nat Struct Mol Biol. 12:110–112. 2005. View Article : Google Scholar : PubMed/NCBI
32	Szyf M: DNA methylation and demethylation as targets for anticancer therapy. Biochemistry (Mosc). 70:533–549. 2005. View Article : Google Scholar
33	Moggs JG, Goodman JI, Trosko JE and Roberts RA: Epigenetics and cancer: Implications for drug discovery and safety assessment. Toxicol Appl Pharmacol. 196:422–430. 2004. View Article : Google Scholar : PubMed/NCBI
34	Snykers S, Vanhaecke T, De Becker A, Papeleu P, Vinken M, Van Riet I and Rogiers V: Chromatin remodeling agent tricho-statin A: A key-factor in the hepatic differentiation of human mesenchymal stem cells derived of adult bone marrow. BMC Dev Biol. 7:242007. View Article : Google Scholar
35	Seo MJ, Suh SY, Bae YC and Jung JS: Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo. Biochem Biophys Res Commun. 328:258–264. 2005. View Article : Google Scholar : PubMed/NCBI
36	Milhem M, Mahmud N, Lavelle D, Araki H, DeSimone J, Saunthararajah Y and Hoffman R: Modification of hematopoietic stem cell fate by 5aza 2′deoxycytidine and trichostatin A. Blood. 103:4102–4110. 2004. View Article : Google Scholar : PubMed/NCBI
37	Sgodda M, Aurich H, Kleist S, Aurich I, König S, Dollinger MM, Fleig WE and Christ B: Hepatocyte differentiation of mesenchymal stem cells from rat peritoneal adipose tissue in vitro and in vivo. Exp Cell Res. 313:2875–2886. 2007. View Article : Google Scholar : PubMed/NCBI
38	Yoshida Y, Shimomura T, Sakabe T, Ishii K, Gonda K, Matsuoka S, Watanabe Y, Takubo K, Tsuchiya H, Hoshikawa Y, et al: A role of Wnt/beta-catenin signals in hepatic fate specification of human umbilical cord blood-derived mesenchymal stem cells. Am J Physiol Gastrointest Liver Physiol. 293:G1089–G1098. 2007. View Article : Google Scholar : PubMed/NCBI
39	Gómez-Lechón MJ, Donato MT, Castell JV and Jover R: Human hepatocytes in primary culture: The choice to investigate drug metabolism in man. Curr Drug Metab. 5:443–462. 2004. View Article : Google Scholar : PubMed/NCBI
40	Kmieć Z: Cooperation of liver cells in health and disease. Adv Anat Embryol Cell Biol. 161:III–XIII. 1–151. 2001.
41	Imamura T, Cui L, Teng R, Johkura K, Okouchi Y, Asanuma K, Ogiwara N and Sasaki K: Embryonic stem cell-derived embryoid bodies in three-dimensional culture system form hepatocyte-like cells in vitro and in vivo. Tissue Eng. 10:1716–1724. 2004. View Article : Google Scholar
42	Liao CX, Tang H, Zhou J, Su J, Zhang SH and Zhang CX: Sieving of hepatic stem cells derived from bone marrow and their potential of differentiation. Chinese Journal of Hepatobiliary Surgery. 9:614–616. 2007.In Chinese.
43	Yuan J, Liao CX, Qin AC, Liao XX, Huang YP, Gong ZY and Liao H: Protocols for cloning human bone marrow-derived hepatic stem cells in vitro. Nan Fang Yi Ke Da Xue Xue Bao. 30:318–320. 2010.In Chinese. PubMed/NCBI
44	Qin AC, Liao CX, Wang Y, Yuan J, Huang YP, Liao XX, Lai YQ and Gong ZY: Intrahepatic transplantation of in vitro induced autologous bone marrow-derived liver stem cells in patients with posthepatitic cirrhosis. Nan Fang Yi Ke Da Xue Xue Bao. 30:529–531. 2010.In Chinese. PubMed/NCBI