Expression of Cancer Stem Cell Markers EpCAM and CD90 Is Correlated with Anti- and Pro-Oncogenic EphA2 Signaling in Hepatocellular Carcinoma
Hepatocellular carcinoma (HCC) is the third leading cause of cancer death worldwide. Additionally, the efficacy of targeted molecular therapies with multiple tyrosine kinase inhibitors is limited. In this study, we focused on the cellular signaling pathways common to diverse HCC cells and used quant...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/16/8652 |
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| author | Nobuhiko Asakura Naotoshi Nakamura Atsushi Muroi Yosui Nojima Taro Yamashita Shuichi Kaneko Kazuki Ikeda Naohiko Koshikawa Takashi Suzuki |
| author_facet | Nobuhiko Asakura Naotoshi Nakamura Atsushi Muroi Yosui Nojima Taro Yamashita Shuichi Kaneko Kazuki Ikeda Naohiko Koshikawa Takashi Suzuki |
| author_sort | Nobuhiko Asakura |
| collection | DOAJ |
| description | Hepatocellular carcinoma (HCC) is the third leading cause of cancer death worldwide. Additionally, the efficacy of targeted molecular therapies with multiple tyrosine kinase inhibitors is limited. In this study, we focused on the cellular signaling pathways common to diverse HCC cells and used quantitative reverse phase protein array (RPPA) and statistical analyses to elucidate the molecular mechanisms determining its malignancy. We examined the heterogeneity of 17 liver cancer cell lines by performing cluster analysis of their expression of CD90 and EpCAM cancer stem cell markers. Gaussian mixture model clustering identified three dominant clusters: CD90-positive and EpCAM-negative (CD90+), EpCAM-positive and CD90-negative (EpCAM+) and EpCAM-negative and CD90-negative (Neutral). A multivariate analysis by partial least squares revealed that the former two cell populations showed distinct patterns of protein expression and phosphorylation in the EGFR and EphA2 signaling pathways. The CD90+ cells exhibited higher abundance of AKT, EphA2 and its phosphorylated form at Ser<sup>897</sup>, whereas the EpCAM+ cells exhibited higher abundance of ERK, RSK and its phosphorylated form. This demonstrates that pro-oncogenic, ligand-independent EphA2 signaling plays a dominant role in CD90+ cells with higher motility and metastatic activity than EpCAM+ cells. We also showed that an AKT inhibitor reduced the proliferation and survival of CD90+ cells but did not affect those of EpCAM+ cells. Taken together, our results suggest that AKT activation may be a key pro-oncogenic regulator in HCC. |
| first_indexed | 2024-03-10T08:44:48Z |
| format | Article |
| id | doaj.art-6ddb411ecd8c43bd9ca62725fac5dbb5 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T08:44:48Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-6ddb411ecd8c43bd9ca62725fac5dbb52023-11-22T07:58:21ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-08-012216865210.3390/ijms22168652Expression of Cancer Stem Cell Markers EpCAM and CD90 Is Correlated with Anti- and Pro-Oncogenic EphA2 Signaling in Hepatocellular CarcinomaNobuhiko Asakura0Naotoshi Nakamura1Atsushi Muroi2Yosui Nojima3Taro Yamashita4Shuichi Kaneko5Kazuki Ikeda6Naohiko Koshikawa7Takashi Suzuki8Center for Mathematical Modeling and Data Science, Osaka University, Osaka 580-8531, JapanCenter for Mathematical Modeling and Data Science, Osaka University, Osaka 580-8531, JapanKanagawa Cancer Center Research Institute, Yokohama 241-8515, JapanCenter for Mathematical Modeling and Data Science, Osaka University, Osaka 580-8531, JapanDepartment of General Medicine, Kanazawa University Hospital, Kanazawa 920-8641, JapanDepartment of Gastroenterology, Kanazawa University Hospital, Kanazawa 920-8641, JapanDepartment of Life Science and Technology, Tokyo Institute of Technology, Yokohama 226-8501, JapanKanagawa Cancer Center Research Institute, Yokohama 241-8515, JapanCenter for Mathematical Modeling and Data Science, Osaka University, Osaka 580-8531, JapanHepatocellular carcinoma (HCC) is the third leading cause of cancer death worldwide. Additionally, the efficacy of targeted molecular therapies with multiple tyrosine kinase inhibitors is limited. In this study, we focused on the cellular signaling pathways common to diverse HCC cells and used quantitative reverse phase protein array (RPPA) and statistical analyses to elucidate the molecular mechanisms determining its malignancy. We examined the heterogeneity of 17 liver cancer cell lines by performing cluster analysis of their expression of CD90 and EpCAM cancer stem cell markers. Gaussian mixture model clustering identified three dominant clusters: CD90-positive and EpCAM-negative (CD90+), EpCAM-positive and CD90-negative (EpCAM+) and EpCAM-negative and CD90-negative (Neutral). A multivariate analysis by partial least squares revealed that the former two cell populations showed distinct patterns of protein expression and phosphorylation in the EGFR and EphA2 signaling pathways. The CD90+ cells exhibited higher abundance of AKT, EphA2 and its phosphorylated form at Ser<sup>897</sup>, whereas the EpCAM+ cells exhibited higher abundance of ERK, RSK and its phosphorylated form. This demonstrates that pro-oncogenic, ligand-independent EphA2 signaling plays a dominant role in CD90+ cells with higher motility and metastatic activity than EpCAM+ cells. We also showed that an AKT inhibitor reduced the proliferation and survival of CD90+ cells but did not affect those of EpCAM+ cells. Taken together, our results suggest that AKT activation may be a key pro-oncogenic regulator in HCC.https://www.mdpi.com/1422-0067/22/16/8652hepatocellular carcinomareverse phase protein arrayliver cancer stem cellsEpCAMCD90EphA2 |
| spellingShingle | Nobuhiko Asakura Naotoshi Nakamura Atsushi Muroi Yosui Nojima Taro Yamashita Shuichi Kaneko Kazuki Ikeda Naohiko Koshikawa Takashi Suzuki Expression of Cancer Stem Cell Markers EpCAM and CD90 Is Correlated with Anti- and Pro-Oncogenic EphA2 Signaling in Hepatocellular Carcinoma International Journal of Molecular Sciences hepatocellular carcinoma reverse phase protein array liver cancer stem cells EpCAM CD90 EphA2 |
| title | Expression of Cancer Stem Cell Markers EpCAM and CD90 Is Correlated with Anti- and Pro-Oncogenic EphA2 Signaling in Hepatocellular Carcinoma |
| title_full | Expression of Cancer Stem Cell Markers EpCAM and CD90 Is Correlated with Anti- and Pro-Oncogenic EphA2 Signaling in Hepatocellular Carcinoma |
| title_fullStr | Expression of Cancer Stem Cell Markers EpCAM and CD90 Is Correlated with Anti- and Pro-Oncogenic EphA2 Signaling in Hepatocellular Carcinoma |
| title_full_unstemmed | Expression of Cancer Stem Cell Markers EpCAM and CD90 Is Correlated with Anti- and Pro-Oncogenic EphA2 Signaling in Hepatocellular Carcinoma |
| title_short | Expression of Cancer Stem Cell Markers EpCAM and CD90 Is Correlated with Anti- and Pro-Oncogenic EphA2 Signaling in Hepatocellular Carcinoma |
| title_sort | expression of cancer stem cell markers epcam and cd90 is correlated with anti and pro oncogenic epha2 signaling in hepatocellular carcinoma |
| topic | hepatocellular carcinoma reverse phase protein array liver cancer stem cells EpCAM CD90 EphA2 |
| url | https://www.mdpi.com/1422-0067/22/16/8652 |
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