Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis
In the present study, the role of a novel protein involved in neurite development and endoplasmic reticulum (ER) stress, canopy homolog 2 (CNPY2), was investigated in mouse and human hepatocarcinogenesis. Firstly, a sensitive quantitative and qualitative detection of protein expression using QSTAR E...
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doaj-f57812fc5376406db522dad3b719e8ed2021-07-23T13:33:56ZengMDPI AGCancers2072-66942021-07-01133613361310.3390/cancers13143613Canopy Homolog 2 as a Novel Molecular Target in HepatocarcinogenesisAnna Kakehashi0Shugo Suzuki1Masayuki Shiota2Nina Raymo3Min Gi4Taro Tachibana5Vasily Stefanov6Hideki Wanibuchi7Department of Molecular Pathology, Osaka City University Graduate School of Medicine, Abeno-ku, 1-4-3 Asahi-machi, Osaka 545-8585, JapanDepartment of Molecular Pathology, Osaka City University Graduate School of Medicine, Abeno-ku, 1-4-3 Asahi-machi, Osaka 545-8585, JapanDepartment of Molecular Biology of Medicine, Osaka City University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka 545-8585, JapanDepartment of Molecular Pathology, Osaka City University Graduate School of Medicine, Abeno-ku, 1-4-3 Asahi-machi, Osaka 545-8585, JapanDepartment of Molecular Pathology, Osaka City University Graduate School of Medicine, Abeno-ku, 1-4-3 Asahi-machi, Osaka 545-8585, JapanDepartment of Bioengineering, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, JapanDepartment of Biochemistry, Faculty of Biology, Saint Petersburg State University, 199034 Saint Petersburg, RussiaDepartment of Molecular Pathology, Osaka City University Graduate School of Medicine, Abeno-ku, 1-4-3 Asahi-machi, Osaka 545-8585, JapanIn the present study, the role of a novel protein involved in neurite development and endoplasmic reticulum (ER) stress, canopy homolog 2 (CNPY2), was investigated in mouse and human hepatocarcinogenesis. Firstly, a sensitive quantitative and qualitative detection of protein expression using QSTAR Elite LC-Ms/Ms was performed for the analysis of lysates of microdissected hepatocellular altered foci (AF), adenomas (HCAs), carcinomas (HCCs) and peri-tumoral livers from C57Bl/6J mice treated with diethylnitrosamine (DEN) and then maintained for 27 or 38 weeks on basal diet. Significant overexpression of 18.5 kDa CNPY2 processed form was demonstrated in AF, HCAs and HCCs, while low expression was observed in the livers of DEN-treated and control mice. Furthermore, CNPY2 elevation in AF and tumors was coordinated with accumulation of numerous cytoskeletal proteins, including cytokeratins 8 and 18, actin, non-muscle myosin and septin 9 and those involved in ER and mitochondrial stresses such as calreticulin, prohibitins 1 and 2 and YME1-like-1. Knockdown of CNPY2 in Huh7 and HepG2 human liver cancer cells resulted in significant suppression of cell survival and invasive potential, inhibition of cyclin D1, induction of p21<sup>Waf1/Cip1</sup> and suppression of the apoptosis inhibitor Bcl2. In contrast, transfection of a mouse CNPY2 (mCNPY2-Ds-Red) vector plasmid in Huh7 and HepG2 cancer cells, with subsequent accumulation of CNPY2 in the ER, resulted in significant increase in cancer cells survival. Clinicopathological analysis in 90 HCV-positive HCC patients, revealed significant association of CNPY2 overexpression with poor overall (<i>p</i> = 0.041) survival. Furthermore, CNPY2 increase was associated with vessel invasion (<i>p</i> = 0.038), poor histological differentiation (<i>p</i> = 0.035) and advanced clinical stage (<i>p</i> = 0.016). In conclusion, CNPY2 is a promising molecular target elevated early in hepatocarcinogenesis and prognostic marker for human HCV-associated HCC. CNPY2 is involved in the processes of ER stress, cell cycle progression, proliferation, survival and invasion of liver tumor cells.https://www.mdpi.com/2072-6694/13/14/3613CNPY2hepatocarcinogenesismicehumanmolecular targetprognostic marker |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anna Kakehashi Shugo Suzuki Masayuki Shiota Nina Raymo Min Gi Taro Tachibana Vasily Stefanov Hideki Wanibuchi |
spellingShingle |
Anna Kakehashi Shugo Suzuki Masayuki Shiota Nina Raymo Min Gi Taro Tachibana Vasily Stefanov Hideki Wanibuchi Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis Cancers CNPY2 hepatocarcinogenesis mice human molecular target prognostic marker |
author_facet |
Anna Kakehashi Shugo Suzuki Masayuki Shiota Nina Raymo Min Gi Taro Tachibana Vasily Stefanov Hideki Wanibuchi |
author_sort |
Anna Kakehashi |
title |
Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis |
title_short |
Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis |
title_full |
Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis |
title_fullStr |
Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis |
title_full_unstemmed |
Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis |
title_sort |
canopy homolog 2 as a novel molecular target in hepatocarcinogenesis |
publisher |
MDPI AG |
series |
Cancers |
issn |
2072-6694 |
publishDate |
2021-07-01 |
description |
In the present study, the role of a novel protein involved in neurite development and endoplasmic reticulum (ER) stress, canopy homolog 2 (CNPY2), was investigated in mouse and human hepatocarcinogenesis. Firstly, a sensitive quantitative and qualitative detection of protein expression using QSTAR Elite LC-Ms/Ms was performed for the analysis of lysates of microdissected hepatocellular altered foci (AF), adenomas (HCAs), carcinomas (HCCs) and peri-tumoral livers from C57Bl/6J mice treated with diethylnitrosamine (DEN) and then maintained for 27 or 38 weeks on basal diet. Significant overexpression of 18.5 kDa CNPY2 processed form was demonstrated in AF, HCAs and HCCs, while low expression was observed in the livers of DEN-treated and control mice. Furthermore, CNPY2 elevation in AF and tumors was coordinated with accumulation of numerous cytoskeletal proteins, including cytokeratins 8 and 18, actin, non-muscle myosin and septin 9 and those involved in ER and mitochondrial stresses such as calreticulin, prohibitins 1 and 2 and YME1-like-1. Knockdown of CNPY2 in Huh7 and HepG2 human liver cancer cells resulted in significant suppression of cell survival and invasive potential, inhibition of cyclin D1, induction of p21<sup>Waf1/Cip1</sup> and suppression of the apoptosis inhibitor Bcl2. In contrast, transfection of a mouse CNPY2 (mCNPY2-Ds-Red) vector plasmid in Huh7 and HepG2 cancer cells, with subsequent accumulation of CNPY2 in the ER, resulted in significant increase in cancer cells survival. Clinicopathological analysis in 90 HCV-positive HCC patients, revealed significant association of CNPY2 overexpression with poor overall (<i>p</i> = 0.041) survival. Furthermore, CNPY2 increase was associated with vessel invasion (<i>p</i> = 0.038), poor histological differentiation (<i>p</i> = 0.035) and advanced clinical stage (<i>p</i> = 0.016). In conclusion, CNPY2 is a promising molecular target elevated early in hepatocarcinogenesis and prognostic marker for human HCV-associated HCC. CNPY2 is involved in the processes of ER stress, cell cycle progression, proliferation, survival and invasion of liver tumor cells. |
topic |
CNPY2 hepatocarcinogenesis mice human molecular target prognostic marker |
url |
https://www.mdpi.com/2072-6694/13/14/3613 |
work_keys_str_mv |
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