Hedgehog Signal Inhibitor GANT61 Inhibits the Malignant Behavior of Undifferentiated Hepatocellular Carcinoma Cells by Targeting Non-Canonical GLI Signaling
The Hedgehog (HH)–GLI pathway plays an important role in cell dedifferentiation and is therefore pivotally involved in the malignant transformation of cancer cells. GANT61, a selective inhibitor of GLI1 and GLI2, was reported as a promising treatment for cancer in various tissues; however, the biolo...
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MDPI AG
2020-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/9/3126 |
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| author | Kensuke Harada Ryuya Ohashi Kyoko Naito Keita Kanki |
| author_facet | Kensuke Harada Ryuya Ohashi Kyoko Naito Keita Kanki |
| author_sort | Kensuke Harada |
| collection | DOAJ |
| description | The Hedgehog (HH)–GLI pathway plays an important role in cell dedifferentiation and is therefore pivotally involved in the malignant transformation of cancer cells. GANT61, a selective inhibitor of GLI1 and GLI2, was reported as a promising treatment for cancer in various tissues; however, the biological impact of GANT61 in hepatocellular carcinoma (HCC), especially in undifferentiated HCC cells, remains unclear. In this study, we investigated the antitumor effect of GANT61 using two undifferentiated hepatoma cell lines: HLE and HLF. Quantitative PCR and RT-PCR analyses revealed that these cells express <i>GLI</i> transcripts, showing mesenchymal phenotypes characterized by the loss of epithelial and hepatic markers and specific expression of epithelial–mesenchymal transition (EMT)-related genes. GANT61 significantly reduced the proliferation and cell viability after drug treatment using 5-FU and Mitomycin C. We showed that <i>GLI</i> transcript levels were down-regulated by the MEK inhibitor U0126 and the Raf inhibitor sorafenib, suggesting that non-canonical signaling including the Ras–Raf–MEK–ERK pathway is involved. Sphere formation and migration were significantly decreased by GANT61 treatment, and it is suggested that the underlying molecular mechanisms are the down-regulation of stemness-related genes (<i>Oct4</i>, <i>Bmi1</i>, <i>CD44,</i> and <i>ALDH</i>) and the EMT-related gene <i>Snail1</i>. The data presented here showed that direct inhibition of GLI might be beneficial for the treatment of dedifferentiated HCC. |
| first_indexed | 2024-03-10T20:10:41Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T20:10:41Z |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9bc1f1a9b14943c29cced6d9f740f2d32023-11-19T22:59:31ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-04-01219312610.3390/ijms21093126Hedgehog Signal Inhibitor GANT61 Inhibits the Malignant Behavior of Undifferentiated Hepatocellular Carcinoma Cells by Targeting Non-Canonical GLI SignalingKensuke Harada0Ryuya Ohashi1Kyoko Naito2Keita Kanki3Department of Biomedical Engineering, Faculty of Engineering, Okayama University of Science, 1-1 Ridai-cho, Kita-ku, Okayama 700-0005, JapanDepartment of Biomedical Engineering, Faculty of Engineering, Okayama University of Science, 1-1 Ridai-cho, Kita-ku, Okayama 700-0005, JapanDepartment of Biomedical Engineering, Faculty of Engineering, Okayama University of Science, 1-1 Ridai-cho, Kita-ku, Okayama 700-0005, JapanDepartment of Biomedical Engineering, Faculty of Engineering, Okayama University of Science, 1-1 Ridai-cho, Kita-ku, Okayama 700-0005, JapanThe Hedgehog (HH)–GLI pathway plays an important role in cell dedifferentiation and is therefore pivotally involved in the malignant transformation of cancer cells. GANT61, a selective inhibitor of GLI1 and GLI2, was reported as a promising treatment for cancer in various tissues; however, the biological impact of GANT61 in hepatocellular carcinoma (HCC), especially in undifferentiated HCC cells, remains unclear. In this study, we investigated the antitumor effect of GANT61 using two undifferentiated hepatoma cell lines: HLE and HLF. Quantitative PCR and RT-PCR analyses revealed that these cells express <i>GLI</i> transcripts, showing mesenchymal phenotypes characterized by the loss of epithelial and hepatic markers and specific expression of epithelial–mesenchymal transition (EMT)-related genes. GANT61 significantly reduced the proliferation and cell viability after drug treatment using 5-FU and Mitomycin C. We showed that <i>GLI</i> transcript levels were down-regulated by the MEK inhibitor U0126 and the Raf inhibitor sorafenib, suggesting that non-canonical signaling including the Ras–Raf–MEK–ERK pathway is involved. Sphere formation and migration were significantly decreased by GANT61 treatment, and it is suggested that the underlying molecular mechanisms are the down-regulation of stemness-related genes (<i>Oct4</i>, <i>Bmi1</i>, <i>CD44,</i> and <i>ALDH</i>) and the EMT-related gene <i>Snail1</i>. The data presented here showed that direct inhibition of GLI might be beneficial for the treatment of dedifferentiated HCC.https://www.mdpi.com/1422-0067/21/9/3126GANT61non-canonical Hedgehog-GLI signalinghepatocellular carcinoma |
| spellingShingle | Kensuke Harada Ryuya Ohashi Kyoko Naito Keita Kanki Hedgehog Signal Inhibitor GANT61 Inhibits the Malignant Behavior of Undifferentiated Hepatocellular Carcinoma Cells by Targeting Non-Canonical GLI Signaling International Journal of Molecular Sciences GANT61 non-canonical Hedgehog-GLI signaling hepatocellular carcinoma |
| title | Hedgehog Signal Inhibitor GANT61 Inhibits the Malignant Behavior of Undifferentiated Hepatocellular Carcinoma Cells by Targeting Non-Canonical GLI Signaling |
| title_full | Hedgehog Signal Inhibitor GANT61 Inhibits the Malignant Behavior of Undifferentiated Hepatocellular Carcinoma Cells by Targeting Non-Canonical GLI Signaling |
| title_fullStr | Hedgehog Signal Inhibitor GANT61 Inhibits the Malignant Behavior of Undifferentiated Hepatocellular Carcinoma Cells by Targeting Non-Canonical GLI Signaling |
| title_full_unstemmed | Hedgehog Signal Inhibitor GANT61 Inhibits the Malignant Behavior of Undifferentiated Hepatocellular Carcinoma Cells by Targeting Non-Canonical GLI Signaling |
| title_short | Hedgehog Signal Inhibitor GANT61 Inhibits the Malignant Behavior of Undifferentiated Hepatocellular Carcinoma Cells by Targeting Non-Canonical GLI Signaling |
| title_sort | hedgehog signal inhibitor gant61 inhibits the malignant behavior of undifferentiated hepatocellular carcinoma cells by targeting non canonical gli signaling |
| topic | GANT61 non-canonical Hedgehog-GLI signaling hepatocellular carcinoma |
| url | https://www.mdpi.com/1422-0067/21/9/3126 |
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