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Enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma
BACKGROUND: Hepatocellular carcinoma (HCC) is among the deadliest cancers due to its heterogeneity, contributing to chemoresistance and recurrence. Cancer stem-like cells (CSCs) are suggested to play an important role in HCC tumorigenesis. This study investigates the role of Wnt/β-catenin pathway in...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6091111/ https://www.ncbi.nlm.nih.gov/pubmed/30075764 http://dx.doi.org/10.1186/s12885-018-4683-0 |
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author | Pandit, Harshul Li, Yan Li, Xuanyi Zhang, Weizhong Li, Suping Martin, Robert C. G. |
author_facet | Pandit, Harshul Li, Yan Li, Xuanyi Zhang, Weizhong Li, Suping Martin, Robert C. G. |
author_sort | Pandit, Harshul |
collection | PubMed |
description | BACKGROUND: Hepatocellular carcinoma (HCC) is among the deadliest cancers due to its heterogeneity, contributing to chemoresistance and recurrence. Cancer stem-like cells (CSCs) are suggested to play an important role in HCC tumorigenesis. This study investigates the role of Wnt/β-catenin pathway in CSC enrichment and the capabilities of these CSCs in tumor initiation in orthotopic immunocompetent mouse model. METHODS: HCC-CSCs were enriched using established serum-free culture method. Wnt/β-catenin pathway activation and its components were analyzed by western blot and qRT-PCR. The role of β-catenin in enrichment of CSC spheroids was confirmed using siRNA interference. Tumorigenic capabilities were confirmed using orthotopic immunocompetent mouse model by injecting 2 × 10(6) Hepa1–6 CSC spheroids or control cells in upper left liver lobe. RESULTS: The serum-free cultured Hepa1–6 cells demonstrated self-renewal, spheroid formation, higher EpCAM expression, increased Hoechst-33342 efflux, and upregulated Wnt/β-catenin signaling. Wnt/β-catenin pathway upregulation was implicated with the downstream targets, i.e., c-MYC, Cyclin-D1, and LEF1. Also, we found that GSK-3β serine-9 phosphorylation increased in Hepa1–6 spheroids. Silencing β-catenin by siRNA reversed spheroid formation phenotype. Mice injected with Hepa1–6 CSC spheroids showed aggressive tumor initiation and growth compared with mice injected with control cells. CONCLUSIONS: Successfully induced Hepa1–6 spheroids were identified with CSC-like properties. Aberrant β-catenin upregulation mediated by GSK-3β was observed in the Hepa1–6 spheroids. The β-catenin mediated CSC enrichment in the induced spheroids possesses the capability of tumor initiation in immunocompetent mice. Our study suggests plausible cell dedifferentiation mediated by β-catenin contributes to CSC-initiated HCC tumor growth in vivo. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12885-018-4683-0) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6091111 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-60911112018-08-20 Enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma Pandit, Harshul Li, Yan Li, Xuanyi Zhang, Weizhong Li, Suping Martin, Robert C. G. BMC Cancer Research Article BACKGROUND: Hepatocellular carcinoma (HCC) is among the deadliest cancers due to its heterogeneity, contributing to chemoresistance and recurrence. Cancer stem-like cells (CSCs) are suggested to play an important role in HCC tumorigenesis. This study investigates the role of Wnt/β-catenin pathway in CSC enrichment and the capabilities of these CSCs in tumor initiation in orthotopic immunocompetent mouse model. METHODS: HCC-CSCs were enriched using established serum-free culture method. Wnt/β-catenin pathway activation and its components were analyzed by western blot and qRT-PCR. The role of β-catenin in enrichment of CSC spheroids was confirmed using siRNA interference. Tumorigenic capabilities were confirmed using orthotopic immunocompetent mouse model by injecting 2 × 10(6) Hepa1–6 CSC spheroids or control cells in upper left liver lobe. RESULTS: The serum-free cultured Hepa1–6 cells demonstrated self-renewal, spheroid formation, higher EpCAM expression, increased Hoechst-33342 efflux, and upregulated Wnt/β-catenin signaling. Wnt/β-catenin pathway upregulation was implicated with the downstream targets, i.e., c-MYC, Cyclin-D1, and LEF1. Also, we found that GSK-3β serine-9 phosphorylation increased in Hepa1–6 spheroids. Silencing β-catenin by siRNA reversed spheroid formation phenotype. Mice injected with Hepa1–6 CSC spheroids showed aggressive tumor initiation and growth compared with mice injected with control cells. CONCLUSIONS: Successfully induced Hepa1–6 spheroids were identified with CSC-like properties. Aberrant β-catenin upregulation mediated by GSK-3β was observed in the Hepa1–6 spheroids. The β-catenin mediated CSC enrichment in the induced spheroids possesses the capability of tumor initiation in immunocompetent mice. Our study suggests plausible cell dedifferentiation mediated by β-catenin contributes to CSC-initiated HCC tumor growth in vivo. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12885-018-4683-0) contains supplementary material, which is available to authorized users. BioMed Central 2018-08-03 /pmc/articles/PMC6091111/ /pubmed/30075764 http://dx.doi.org/10.1186/s12885-018-4683-0 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Pandit, Harshul Li, Yan Li, Xuanyi Zhang, Weizhong Li, Suping Martin, Robert C. G. Enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma |
title | Enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma |
title_full | Enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma |
title_fullStr | Enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma |
title_full_unstemmed | Enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma |
title_short | Enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma |
title_sort | enrichment of cancer stem cells via β-catenin contributing to the tumorigenesis of hepatocellular carcinoma |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6091111/ https://www.ncbi.nlm.nih.gov/pubmed/30075764 http://dx.doi.org/10.1186/s12885-018-4683-0 |
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