AXL Knock-Out in SNU475 Hepatocellular Carcinoma Cells Provides Evidence for Lethal Effect Associated with G2 Arrest and Polyploidization
AXL, a member of the TAM family, is a promising therapeutic target due to its elevated expression in advanced hepatocellular carcinoma (HCC), particularly in association with acquired drug resistance. Previously, RNA interference was used to study its role in cancer, and several phenotypic changes,...
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MDPI AG
2021-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/24/13247 |
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| author | Tugce Batur Ayse Argundogan Umur Keles Zeynep Mutlu Hani Alotaibi Serif Senturk Mehmet Ozturk |
| author_facet | Tugce Batur Ayse Argundogan Umur Keles Zeynep Mutlu Hani Alotaibi Serif Senturk Mehmet Ozturk |
| author_sort | Tugce Batur |
| collection | DOAJ |
| description | AXL, a member of the TAM family, is a promising therapeutic target due to its elevated expression in advanced hepatocellular carcinoma (HCC), particularly in association with acquired drug resistance. Previously, RNA interference was used to study its role in cancer, and several phenotypic changes, including attenuated cell proliferation and decreased migration and invasion, have been reported. The mechanism of action of AXL in HCC is elusive. We first studied the AXL expression in HCC cell lines by real-time PCR and western blot and showed its stringent association with a mesenchymal phenotype. We then explored the role of AXL in mesenchymal SNU475 cells by CRISPR-Cas9 mediated gene knock-out. AXL-depleted HCC cells displayed drastic phenotypic changes, including increased DNA damage response, prolongation of doubling time, G2 arrest, and polyploidization in vitro and loss of tumorigenicity in vivo. Pharmacological inhibition of AXL by R428 recapitulated G2 arrest and polyploidy phenotype. These observations strongly suggest that acute loss of AXL in some mesenchymal HCC cells is lethal and points out that its inhibition may represent a druggable vulnerability in AXL-high HCC patients. |
| first_indexed | 2024-03-10T03:57:07Z |
| format | Article |
| id | doaj.art-fc67562b41124bd597d358471f2540be |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:57:07Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-fc67562b41124bd597d358471f2540be2023-11-23T08:43:27ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-12-0122241324710.3390/ijms222413247AXL Knock-Out in SNU475 Hepatocellular Carcinoma Cells Provides Evidence for Lethal Effect Associated with G2 Arrest and PolyploidizationTugce Batur0Ayse Argundogan1Umur Keles2Zeynep Mutlu3Hani Alotaibi4Serif Senturk5Mehmet Ozturk6Izmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Izmir 35330, TurkeyIzmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Izmir 35330, TurkeyIzmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Izmir 35330, TurkeyIzmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Izmir 35330, TurkeyIzmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Izmir 35330, TurkeyIzmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Izmir 35330, TurkeyIzmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Izmir 35330, TurkeyAXL, a member of the TAM family, is a promising therapeutic target due to its elevated expression in advanced hepatocellular carcinoma (HCC), particularly in association with acquired drug resistance. Previously, RNA interference was used to study its role in cancer, and several phenotypic changes, including attenuated cell proliferation and decreased migration and invasion, have been reported. The mechanism of action of AXL in HCC is elusive. We first studied the AXL expression in HCC cell lines by real-time PCR and western blot and showed its stringent association with a mesenchymal phenotype. We then explored the role of AXL in mesenchymal SNU475 cells by CRISPR-Cas9 mediated gene knock-out. AXL-depleted HCC cells displayed drastic phenotypic changes, including increased DNA damage response, prolongation of doubling time, G2 arrest, and polyploidization in vitro and loss of tumorigenicity in vivo. Pharmacological inhibition of AXL by R428 recapitulated G2 arrest and polyploidy phenotype. These observations strongly suggest that acute loss of AXL in some mesenchymal HCC cells is lethal and points out that its inhibition may represent a druggable vulnerability in AXL-high HCC patients.https://www.mdpi.com/1422-0067/22/24/13247AXLhepatocellular carcinomaCRISPR-Cas9gene knock-outcell cycle arrestDNA damage |
| spellingShingle | Tugce Batur Ayse Argundogan Umur Keles Zeynep Mutlu Hani Alotaibi Serif Senturk Mehmet Ozturk AXL Knock-Out in SNU475 Hepatocellular Carcinoma Cells Provides Evidence for Lethal Effect Associated with G2 Arrest and Polyploidization International Journal of Molecular Sciences AXL hepatocellular carcinoma CRISPR-Cas9 gene knock-out cell cycle arrest DNA damage |
| title | AXL Knock-Out in SNU475 Hepatocellular Carcinoma Cells Provides Evidence for Lethal Effect Associated with G2 Arrest and Polyploidization |
| title_full | AXL Knock-Out in SNU475 Hepatocellular Carcinoma Cells Provides Evidence for Lethal Effect Associated with G2 Arrest and Polyploidization |
| title_fullStr | AXL Knock-Out in SNU475 Hepatocellular Carcinoma Cells Provides Evidence for Lethal Effect Associated with G2 Arrest and Polyploidization |
| title_full_unstemmed | AXL Knock-Out in SNU475 Hepatocellular Carcinoma Cells Provides Evidence for Lethal Effect Associated with G2 Arrest and Polyploidization |
| title_short | AXL Knock-Out in SNU475 Hepatocellular Carcinoma Cells Provides Evidence for Lethal Effect Associated with G2 Arrest and Polyploidization |
| title_sort | axl knock out in snu475 hepatocellular carcinoma cells provides evidence for lethal effect associated with g2 arrest and polyploidization |
| topic | AXL hepatocellular carcinoma CRISPR-Cas9 gene knock-out cell cycle arrest DNA damage |
| url | https://www.mdpi.com/1422-0067/22/24/13247 |
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