{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,17]],"date-time":"2026-04-17T11:10:44Z","timestamp":1776424244125,"version":"3.51.2"},"reference-count":24,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T00:00:00Z","timestamp":1775692800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mixed hepatocellular carcinoma (HCC)\u2013neuroendocrine carcinoma (NEC) is a major type of liver mixed neuroendocrine\u2013non-neuroendocrine neoplasm (MiNEN). Primary liver NEC, which is very rare, is mostly associated with HCC rather than pure NEC. To characterize the cancer cell heterogeneity of the HCC and NEC components, we comprehensively analyzed the protein expression of three cancer cell biological markers (TERT, Ki-67, and p53) and five differentiation markers (one hepatocyte marker and four neuroendocrine markers) via immunohistochemistry and immunofluorescence using curative resection tissues from three patients with liver MiNEN. TERT\/Ki-67\/p53 proteins, which are related to cell proliferation and malignancy, were independently expressed in the HCC and NEC components; Ki-67 was highly expressed among the three proteins in both cancer components, and the expression of all three markers was higher in the NEC component than in the HCC component. Despite the intracomponent and intercomponent heterogeneity, the expression signatures of the three markers were similar between the two components, potentially suggesting a common origin of mixed HCC-NEC. An in-depth exploration of intracomponent heterogeneity using differentiation markers revealed multiple intermediate phenotypes of cancer cells, i.e., HCC-like and NEC-like cells, mainly in the HCC component. Histologically NEC-like cells rather than HCC-like cells tended to have an intermediate percentage of Ki-67-positive cells, compared with NEC cells. The spatial distribution of various intermediate cancer cell phenotypes suggests that mixed HCC-NEC may involve the transdifferentiation from HCC cells to NEC cells through the dedifferentiation of HCC.<\/jats:p>","DOI":"10.3390\/ijms27083390","type":"journal-article","created":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T07:55:12Z","timestamp":1775807712000},"page":"3390","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Heterogeneous Intermediate Phenotypes of Cancer Cells with Varying Ki-67-Positivity Rates, Including Histologically HCC-like and NEC-like Cells, in Liver MiNEN"],"prefix":"10.3390","volume":"27","author":[{"given":"Sumie","family":"Ohni","sequence":"first","affiliation":[{"name":"Division of Oncologic Pathology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoko","family":"Nakanishi","sequence":"additional","affiliation":[{"name":"Division of Oncologic Pathology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yukari","family":"Hirotani","sequence":"additional","affiliation":[{"name":"Division of Oncologic Pathology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryosuke","family":"Toyonaka","sequence":"additional","affiliation":[{"name":"Division of Digestive Surgery, Department of Surgery, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4264-4384","authenticated-orcid":false,"given":"Osamu","family":"Aramaki","sequence":"additional","affiliation":[{"name":"Division of Digestive Surgery, Department of Surgery, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yukiyasu","family":"Okamura","sequence":"additional","affiliation":[{"name":"Division of Digestive Surgery, Department of Surgery, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shinobu","family":"Masuda","sequence":"additional","affiliation":[{"name":"Division of Oncologic Pathology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4630-905X","authenticated-orcid":false,"given":"Makoto","family":"Makishima","sequence":"additional","affiliation":[{"name":"Division of Biochemistry, Department of Biomedical Sciences, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4253-2027","authenticated-orcid":false,"given":"Mariko","family":"Esumi","sequence":"additional","affiliation":[{"name":"Division of Biochemistry, Department of Biomedical Sciences, Nihon University School of Medicine, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,9]]},"reference":[{"key":"ref_1","unstructured":"WHO Classification of Tumours Editorial Board (2019). Classification of neuroendocrine neoplasms of the digestive system. Who Classification of Tumours, Digestive System Tumours, IARC. [5th ed.]."},{"key":"ref_2","unstructured":"WHO Classification of Tumours Editorial Board (2019). Hepatic neuroendocrine neoplasms. WHO Classification of Tumours, Digestive System Tumours, IARC. [5th ed.]."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Gao, X., Wang, H., Niu, Z., Liu, M., Kong, X., Sun, H., Ma, C., Zhu, H., Lu, J., and Zhou, X. (2024). Case report: Mixed large-cell neuroendocrine and hepatocellular carcinoma of the liver. Front. Oncol., 13.","DOI":"10.3389\/fonc.2023.1309798"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Sambataro, D., Bellavia, S., Di Mattia, P., Centonze, D., Emmanuele, C., Bonasera, A., Caputo, G., Quattrocchi, A.M.O., Vinci, E., and Gebbia, V. (2025). Combined neuroendocrine carcinoma and hepatocellular carcinoma of the liver: Systematic literature review suggests implementing biological characterization to optimize therapeutic strategy. Cancers, 17.","DOI":"10.3390\/cancers17071074"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"690","DOI":"10.62347\/CQJW7490","article-title":"Complex phenotypic heterogeneity of combined hepatocellular-cholangiocarcinoma with a homogenous TERT promoter mutation","volume":"16","author":"Ohni","year":"2024","journal-title":"Am. J. Transl. Res."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Liu, J., Li, W., Deng, M., Liu, D., Ma, Q., and Feng, X. (2016). Immunohistochemical determination of p53 protein overexpression for predicting p53 gene mutations in hepatocellular carcinoma: A meta-analysis. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0159636"},{"key":"ref_7","first-page":"10235","article-title":"Clinicopathological and prognostic significance of high ki-67 labeling index in hepatocellular carcinoma patients: A meta-analysis","volume":"8","author":"Luo","year":"2015","journal-title":"Int. J. Clin. Exp. Med."},{"key":"ref_8","first-page":"21","article-title":"Mixed neuroendocrine non-neuroendocrine tumor (minen) of the liver: Report of two cases and review of the literature","volume":"41","author":"Mubeen","year":"2025","journal-title":"Turk. J. Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.humpath.2021.07.007","article-title":"Primary hepatic neoplasms arising in cirrhotic livers can have a variable spectrum of neuroendocrine differentiation","volume":"116","author":"Shi","year":"2021","journal-title":"Hum. Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1186\/s12957-021-02187-5","article-title":"Combined primary hepatic neuroendocrine carcinoma and hepatocellular carcinoma: Case report and literature review","volume":"19","author":"Nakano","year":"2021","journal-title":"World J. Surg. Oncol."},{"key":"ref_11","first-page":"100020","article-title":"Mixed hepatocellular carcinoma-neuroendocrine carcinoma\u2014A diagnostic and therapeutic challenge","volume":"1","author":"Jahan","year":"2020","journal-title":"Curr. Probl. Cancer Case Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1136\/jclinpath-2016-203941","article-title":"Clinicopathological features of neoplasms with neuroendocrine differentiation occurring in the liver","volume":"70","author":"Nomura","year":"2017","journal-title":"J. Clin. Pathol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1007\/s00428-006-0251-0","article-title":"Tumor of the liver (hepatocellular and high grade neuroendocrine carcinoma): A case report and review of the literature","volume":"449","author":"Garcia","year":"2006","journal-title":"Virchows Arch."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1111\/j.1440-1827.2004.01770.x","article-title":"Hepatocellular carcinoma with an unusual neuroendocrine component","volume":"54","author":"Yamaguchi","year":"2004","journal-title":"Pathol. Int."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1121","DOI":"10.1177\/000313481608201130","article-title":"Mixed hepatocellular carcinoma, neuroendocrine carcinoma of the liver","volume":"82","author":"Baker","year":"2016","journal-title":"Am. Surg."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5620","DOI":"10.1016\/j.cell.2023.11.006","article-title":"Molecular cartography uncovers evolutionary and microenvironmental dynamics in sporadic colorectal tumors","volume":"186","author":"Heiser","year":"2023","journal-title":"Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1016\/j.ccell.2024.05.002","article-title":"Molecular subtypes of neuroendocrine carcinomas: A cross-tissue classification framework based on five transcriptional regulators","volume":"42","author":"Wang","year":"2024","journal-title":"Cancer Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"8290","DOI":"10.1038\/s41467-023-43749-3","article-title":"Deep learning-based phenotyping reclassifies combined hepatocellular-cholangiocarcinoma","volume":"14","author":"Calderaro","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Lan, J., Guo, D., Qin, X., Chen, B., and Liu, Q. (2021). Mixed neuroendocrine carcinoma and hepatocellular carcinoma: A case report and literature review. Front. Surg., 8.","DOI":"10.3389\/fsurg.2021.678853"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2342","DOI":"10.1016\/j.ajpath.2015.06.004","article-title":"Hepatic progenitor cells in action: Liver regeneration or fibrosis?","volume":"185","author":"Kaur","year":"2015","journal-title":"Am. J. Pathol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/j.ajpath.2023.11.013","article-title":"Clonal origin and lineage ambiguity in mixed neuroendocrine carcinoma of the uterine cervix","volume":"194","author":"Masuda","year":"2024","journal-title":"Am. J. Pathol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1038\/s41586-025-09503-z","article-title":"Basal cell of origin resolves neuroendocrine-tuft lineage plasticity in cancer","volume":"647","author":"Ireland","year":"2025","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1002\/1097-0142(195405)7:3<462::AID-CNCR2820070308>3.0.CO;2-E","article-title":"Primary carcinoma of the liver: A study of 100 cases among 48,900 necropsies","volume":"7","author":"Edmondson","year":"1954","journal-title":"Cancer"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/S0928-4346(96)00325-8","article-title":"New inuyama classification; new criteria for histological assessment of chronic hepatitis","volume":"6","author":"Ichida","year":"1996","journal-title":"Int. Hepatol. 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