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http://dx.doi.org/10.5483/BMBRep.2022.55.8.071

Ahnak depletion accelerates liver regeneration by modulating the TGF-β/Smad signaling pathway  

Yang, Insook (Laboratory of Developmental Biology and Genomics, College of Veterinary Medicine, Seoul National University)
Son, Yeri (Laboratory of Developmental Biology and Genomics, College of Veterinary Medicine, Seoul National University)
Shin, Jae Hoon (Department of Surgery, University of Michigan)
Kim, Il Yong (Korea Mouse Phenotyping Center, Seoul National University)
Seong, Je Kyung (Laboratory of Developmental Biology and Genomics, College of Veterinary Medicine, Seoul National University)
Publication Information
BMB Reports / v.55, no.8, 2022 , pp. 401-406 More about this Journal
Abstract
Ahnak, a large protein first identified as an inhibitor of TGF-β signaling in human neuroblastoma, was recently shown to promote TGF-β in some cancers. The TGF-β signaling pathway regulates cell growth, various biological functions, and cancer growth and metastasis. In this study, we used Ahnak knockout (KO) mice that underwent a 70% partial hepatectomy (PH) to investigate the function of Ahnak in TGF-β signaling during liver regeneration. At the indicated time points after PH, we analyzed the mRNA and protein expression of the TGF -β/Smad signaling pathway and cell cycle-related factors, evaluated the cell cycle through proliferating cell nuclear antigen (PCNA) immunostaining, analyzed the mitotic index by hematoxylin and eosin staining. We also measured the ratio of liver tissue weight to body weight. Activation of TGF-β signaling was confirmed by analyzing the levels of phospho-Smad 2 and 3 in the liver at the indicated time points after PH and was lower in Ahnak KO mice than in WT mice. The expression levels of cyclin B1, D1, and E1; proteins in the Rb/E2F transcriptional pathway, which regulates the cell cycle; and the numbers of PCNA-positive cells were increased in Ahnak KO mice and showed tendencies opposite that of TGF-β expression. During postoperative regeneration, the liver weight to body weight ratio tended to increase faster in Ahnak KO mice. However, 7 days after PH, both groups of mice showed similar rates of regeneration, following which their active regeneration stopped. Analysis of hepatocytes undergoing mitosis showed that there were more mitotic cells in Ahnak KO mice, consistent with the weight ratio. Our findings suggest that Ahnak enhances TGF-β signaling during postoperative liver regeneration, resulting in cell cycle disruption; this highlights a novel role of Ahnak in liver regeneration. These results provide new insight into liver regeneration and potential treatment targets for liver diseases that require surgical treatment.
Keywords
70% partial hepatectomy; Ahnak KO mice; Hepatocyte proliferation; Liver regeneration; $TGF-{\beta}$/Smad signaling pathway;
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1 Fausto N, Campbell JS and Riehle KJ (2006) Liver regeneration. Hepatology 43, S45-53   DOI
2 Michalopoulos GK (2017) Hepatostat: liver regeneration and normal liver tissue maintenance. Hepatology 65, 1384-1392   DOI
3 Kitamura T, Watanabe S and Sato N (1998) Liver regeneration, liver cancers and cyclins. J Gastroenterol Hepatol 13, S96-S99   DOI
4 LK M (2008) Distinct proliferative and transcriptional effects of the D-type cyclins in vivo. Cell Cycle 7, 2215-2224   DOI
5 Liu HX, Fang Y, Hu Y, Gonzalez FJ, Fang J and Wan YJ (2013) PPARbeta regulates liver regeneration by modulating Akt and E2f signaling. PLoS One 8, e65644
6 Assy N, Gong Y, Zhang M, Pettigrew NM, Pashniak D and Minuk GY (1998) Use of proliferating cell nuclear antigen as a marker of liver regeneration after partial hepatectomy in rats. J Lab Clin Med 131, 251-256   DOI
7 Michalopoulos GK and Bhushan B (2021) Liver regeneration: biological and pathological mechanisms and implications. Nat Rev Gastroenterol Hepatol 18, 40-55   DOI
8 Webber EM, Bruix J, Pierce RH and Fausto N (1998) Tumor necrosis factor primes hepatocytes for DNA replication in the rat. Hepatology 28, 1226-1234   DOI
9 Jakowlew SB, Mead JE, Danielpour D, Wu J, Roberts AB and Fausto N (1991) Transforming growth factor-beta (TGF-beta) isoforms in rat liver regeneration: messenger RNA expression and activation of latent TGF-beta. Cell Regul 2, 535-548   DOI
10 Bottinger EP, Factor VM, Tsang ML et al (1996) The recombinant proregion of transforming growth factor beta1 (latency-associated peptide) inhibits active transforming growth factor beta1 in transgenic mice. Proc Natl Acad Sci U S A 93, 5877-5882   DOI
11 Higgins GM (1931) Experimental pathology of the liver. I. Restoration of the liver of the white rat following partial surgical removal. Arch Pathol 12, 186-202
12 Mitchell C and Willenbring H (2008) A reproducible and well-tolerated method for 2/3 partial hepatectomy in mice. Nat Protoc 3, 1167-1170   DOI
13 Michalopoulos GK (2007) Liver regeneration. J Cell Physiol 213, 286-300   DOI
14 Kokura K, Kim H, Shinagawa T, Khan MM, Nomura T and Ishii S (2003) The Ski-binding protein C184M negatively regulates tumor growth factor-beta signaling by sequestering the Smad proteins in the cytoplasm. J Biol Chem 278, 20133-20139   DOI
15 Michalopoulos GK (2010) Liver regeneration after partial hepatectomy: critical analysis of mechanistic dilemmas. Am J Pathol 176, 2-13   DOI
16 Albrecht JH, Poon RY, Ahonen CL, Rieland BM, Deng C and Crary GS (1998) Involvement of p21 and p27 in the regulation of CDK activity and cell cycle progression in the regenerating liver. Oncogene 16, 2141-2150   DOI
17 Shtivelman E, Cohen FE and Bishop JM (1992) A human gene (AHNAK) encoding an unusually large protein with a 1.2-microns polyionic rod structure. Proc Natl Acad Sci U S A 89, 5472-5476   DOI
18 Han H and Kursula P (2014) Periaxin and AHNAK nucleoprotein 2 form intertwined homodimers through domain swapping. J Biol Chem 289, 14121-14131   DOI
19 Dempsey BR, Rezvanpour A, Lee TW, Barber KR, Junop MS and Shaw GS (2012) Structure of an asymmetric ternary protein complex provides insight for membrane interaction. Structure 20, 1737-1745   DOI
20 Peng R, Zhang PF, Yang X et al (2019) Overexpression of RNF38 facilitates TGF-beta signaling by Ubiquitinating and degrading AHNAK in hepatocellular carcinoma. J Exp Clin Cancer Res 38, 113
21 Fausto N (2000) Liver regeneration. J Hepatol 32, 19-31   DOI
22 Park JW, Kim IY, Choi JW et al (2018) AHNAK loss in mice promotes type II pneumocyte hyperplasia and lung tumor development. Mol Cancer Res 16, 1287-1298   DOI
23 Romero-Gallo J, Sozmen EG, Chytil A et al (2005) Inactivation of TGF-beta signaling in hepatocytes results in an increased proliferative response after partial hepatectomy. Oncogene 24, 3028-3041   DOI
24 Baak JP (1990) Mitosis counting in tumors. Hum Pathol 21, 683-685   DOI
25 Oe S, Lemmer ER, Conner EA et al (2004) Intact signaling by transforming growth factor beta is not required for termination of liver regeneration in mice. Hepatology 40, 1098-1105   DOI
26 Nevins JR (2001) The Rb/E2F pathway and cancer. Hum Mol Genet 10, 699-703   DOI
27 Goggin MM, Nelsen CJ, Kimball SR, Jefferson LS, Morley SJ and Albrecht JH (2004) Rapamycin-sensitive induction of eukaryotic initiation factor 4F in regenerating mouse liver. Hepatology 40, 537-544   DOI
28 Kogure K, Zhang YQ, Maeshima A, Suzuki K, Kuwano H and Kojima I (2000) The role of activin and transforming growth factor-beta in the regulation of organ mass in the rat liver. Hepatology 31, 916-921   DOI
29 Wrighton KH, Lin X and Feng XH (2009) Phospho-control of TGF-beta superfamily signaling. Cell Res 19, 8-20   DOI
30 Moriuchi A, Hirono S, Ido A et al (2001) Additive and inhibitory effects of simultaneous treatment with growth factors on DNA synthesis through MAPK pathway and G1 cyclins in rat hepatocytes. Biochem Biophys Res Commun 280, 368-373   DOI
31 Marongiu F, Marongiu M, Contini A et al (2017) Hyperplasia vs hypertrophy in tissue regeneration after extensive liver resection. World J Gastroenterol 23, 1764-1770   DOI
32 Lee IH, Lim HJ, Yoon S et al (2008) Ahnak protein activates protein kinase C (PKC) through dissociation of the PKC-protein phosphatase 2A complex. J Biol Chem 283, 6312-6320   DOI
33 Taub R (2004) Liver regeneration: from myth to mechanism. Nat Rev Mol Cell Biol 5, 836-847   DOI
34 Shankar J, Messenberg A, Chan J, Underhill TM, Foster LJ and Nabi IR (2010) Pseudopodial actin dynamics control epithelial-mesenchymal transition in metastatic cancer cells. Cancer Res 70, 3780-3790
35 Lee IH, Sohn M, Lim HJ et al (2014) Ahnak functions as a tumor suppressor via modulation of TGFbeta/Smad signaling pathway. Oncogene 33, 4675-4684   DOI