USP7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of ARF4
Abstract Background Glioblastomas (GBMs) are grade IV central nervous system tumors characterized by a poor prognosis and a short median overall survival. Effective induction of GBM cell death is difficult because the GBM cell population is genetically unstable, resistant to chemotherapy and highly...
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| Format: | Article |
| Language: | English |
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BMC
2021-09-01
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| Series: | Cancer Cell International |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12935-021-02208-z |
| _version_ | 1819111008217595904 |
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| author | Tingzheng Pan Xuetao Li Yanyan Li Zhennan Tao Hui Yao Yue Wu Guangliang Chen Kai Zhang Youxin Zhou Yulun Huang |
| author_facet | Tingzheng Pan Xuetao Li Yanyan Li Zhennan Tao Hui Yao Yue Wu Guangliang Chen Kai Zhang Youxin Zhou Yulun Huang |
| author_sort | Tingzheng Pan |
| collection | DOAJ |
| description | Abstract Background Glioblastomas (GBMs) are grade IV central nervous system tumors characterized by a poor prognosis and a short median overall survival. Effective induction of GBM cell death is difficult because the GBM cell population is genetically unstable, resistant to chemotherapy and highly angiogenic. In recent studies, ubiquitin-specific protease 7 (USP7) is shown to scavenge ubiquitin from oncogenic protein substrates, so effective inhibition of USP7 may be a potential key treatment for GBM. Methods Immunohistochemistry and western blotting were used to detect the expression of USP7 in GBM tissues. In vitro apoptosis assay of USP7 inhibition was performed by western blotting, immunofluorescence, and flow cytometry. Anti-apoptotic substrates of USP7 were defined by Co-IP and TMT proteomics. Western blotting and IP were used to verify the relationship between USP7 and its substrate. In an in vivo experiment using an intracranial xenograft model in nude mice was constructed to assess the therapeutic effect of target USP7. Results Immunohistochemistry and western blotting confirmed that USP7 was significantly upregulated in glioblastoma samples. In in vitro experiments, inhibition of USP7 in GBM induced significant apoptosis. Co-IP and TMT proteomics identified a key anti-apoptotic substrate of USP7, ADP-ribosylation factor 4 (ARF4). Western blotting and IP confirmed that USP7 interacted directly with ARF4 and catalyzed the removal of the K48-linked polyubiquitinated chain that binded to ARF4. In addition, in vivo experiments revealed that USP7 inhibition significantly suppressed tumor growth and promoted the expression of apoptotic genes. Conclusions Targeted inhibition of USP7 enhances the ubiquitination of ARF4 and ultimately mediates the apoptosis of GBM cells. In a clinical sense, P5091 as a novel specific inhibitor of USP7 may be an effective approach for the treatment of GBM. |
| first_indexed | 2024-12-22T03:50:47Z |
| format | Article |
| id | doaj.art-5a28170dc96147c6a38143d69d98f12f |
| institution | Directory Open Access Journal |
| issn | 1475-2867 |
| language | English |
| last_indexed | 2024-12-22T03:50:47Z |
| publishDate | 2021-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer Cell International |
| spelling | doaj.art-5a28170dc96147c6a38143d69d98f12f2022-12-21T18:40:00ZengBMCCancer Cell International1475-28672021-09-0121111510.1186/s12935-021-02208-zUSP7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of ARF4Tingzheng Pan0Xuetao Li1Yanyan Li2Zhennan Tao3Hui Yao4Yue Wu5Guangliang Chen6Kai Zhang7Youxin Zhou8Yulun Huang9Neurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityDepartment of Neurosurgery, Dushu Lake Hospital Affiliated of Soochow UniversityNeurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityNeurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityNeurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityNeurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityNeurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityNeurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityNeurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityNeurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow UniversityAbstract Background Glioblastomas (GBMs) are grade IV central nervous system tumors characterized by a poor prognosis and a short median overall survival. Effective induction of GBM cell death is difficult because the GBM cell population is genetically unstable, resistant to chemotherapy and highly angiogenic. In recent studies, ubiquitin-specific protease 7 (USP7) is shown to scavenge ubiquitin from oncogenic protein substrates, so effective inhibition of USP7 may be a potential key treatment for GBM. Methods Immunohistochemistry and western blotting were used to detect the expression of USP7 in GBM tissues. In vitro apoptosis assay of USP7 inhibition was performed by western blotting, immunofluorescence, and flow cytometry. Anti-apoptotic substrates of USP7 were defined by Co-IP and TMT proteomics. Western blotting and IP were used to verify the relationship between USP7 and its substrate. In an in vivo experiment using an intracranial xenograft model in nude mice was constructed to assess the therapeutic effect of target USP7. Results Immunohistochemistry and western blotting confirmed that USP7 was significantly upregulated in glioblastoma samples. In in vitro experiments, inhibition of USP7 in GBM induced significant apoptosis. Co-IP and TMT proteomics identified a key anti-apoptotic substrate of USP7, ADP-ribosylation factor 4 (ARF4). Western blotting and IP confirmed that USP7 interacted directly with ARF4 and catalyzed the removal of the K48-linked polyubiquitinated chain that binded to ARF4. In addition, in vivo experiments revealed that USP7 inhibition significantly suppressed tumor growth and promoted the expression of apoptotic genes. Conclusions Targeted inhibition of USP7 enhances the ubiquitination of ARF4 and ultimately mediates the apoptosis of GBM cells. In a clinical sense, P5091 as a novel specific inhibitor of USP7 may be an effective approach for the treatment of GBM.https://doi.org/10.1186/s12935-021-02208-zGBMUSP7UbiquitinationARF4Apoptosis |
| spellingShingle | Tingzheng Pan Xuetao Li Yanyan Li Zhennan Tao Hui Yao Yue Wu Guangliang Chen Kai Zhang Youxin Zhou Yulun Huang USP7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of ARF4 Cancer Cell International GBM USP7 Ubiquitination ARF4 Apoptosis |
| title | USP7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of ARF4 |
| title_full | USP7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of ARF4 |
| title_fullStr | USP7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of ARF4 |
| title_full_unstemmed | USP7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of ARF4 |
| title_short | USP7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of ARF4 |
| title_sort | usp7 inhibition induces apoptosis in glioblastoma by enhancing ubiquitination of arf4 |
| topic | GBM USP7 Ubiquitination ARF4 Apoptosis |
| url | https://doi.org/10.1186/s12935-021-02208-z |
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