RNF8 is responsible for ATRA resistance in variant acute promyelocytic leukemia with GTF2I/RARA fusion, and inhibition of the ubiquitin–proteasome pathway contributes to the reversion of ATRA resistance
Abstract Background GTF2I-RARA is a newly identified RARA fusion gene in variant acute promyelocytic leukemia (APL) patients with t(7;17)(q11;q21). Clinical manifestation in the patient showed that it is a sort of ATRA-insensitive oncogene and is different from the classic PML-RARA in terms of thera...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-04-01
|
| Series: | Cancer Cell International |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12935-019-0803-4 |
| _version_ | 1819168229546786816 |
|---|---|
| author | Wenzhe Yan Ji Li Yang Zhang Yafei Yin Zhao Cheng Jiayi Wang Guoyu Hu Sufang Liu Yewei Wang Yunxiao Xu Hongling Peng Guangsen Zhang |
| author_facet | Wenzhe Yan Ji Li Yang Zhang Yafei Yin Zhao Cheng Jiayi Wang Guoyu Hu Sufang Liu Yewei Wang Yunxiao Xu Hongling Peng Guangsen Zhang |
| author_sort | Wenzhe Yan |
| collection | DOAJ |
| description | Abstract Background GTF2I-RARA is a newly identified RARA fusion gene in variant acute promyelocytic leukemia (APL) patients with t(7;17)(q11;q21). Clinical manifestation in the patient showed that it is a sort of ATRA-insensitive oncogene and is different from the classic PML-RARA in terms of therapeutic reaction. Methods To reveal the functional characteristics and regulating mechanism of the GTF2I-RARA fusion gene, we established a GTF2I-RARA-transfected HL60 cell model and examined its sensitivity to ATRA by western blot, MTT assay, flow cytometry, and Wright-Giemsa staining. Coimmunoprecipitation and confocal microscopy were used to examine the binding of GTF2I-RARA and transcriptional corepressors. We also performed ChIP-seq to search for potential target genes. Immunoprecipitation, ubiquitination assay, western blot, luciferase assay, and real-time PCR were used to analyze the effects of RNF8 on RARA. Flow cytometry and Wright-Giemsa staining were used to study the effect of MG132 and ATRA on the GTF2I-RARA-transfected HL60 cell model. Result We confirmed resistance of GTF2I-RARA to ATRA. Compared with PML-RARA, GTF2I-RARA has a higher affinity to HDAC3 under ATRA treatment. Using the ChIP-sequencing approach, we identified 221 GTF2I-RARA binding sites in model cells and found that the RING finger protein 8 (RNF8) is a target gene of GTF2I-RARA. RNF8 participates in disease progression and therapy resistance in APL with the GTF2I-RARA transcript. Elevated RNF8 expression promotes the interaction between RARA and RNF8 and induces RARA Lys-48 linkage ubiquitylation and degradation, resulting in attenuated transcriptional activation of RARA. Conclusion Our results suggest that RNF8 is a key GTF2I-RARA downstream event. Using the combination of MG132 and ATRA to treat GTF2I-RARA-HL60 cells, a synergistic effect leading to GTF2I-RARA-HL60 cell differentiation was confirmed. Taken together, the targeting of RNF8 may be an alternative choice for treatment in variant APL with GTF2I-RARA fusion. |
| first_indexed | 2024-12-22T19:00:17Z |
| format | Article |
| id | doaj.art-aae3d337e416402da1cb6a4febe231ec |
| institution | Directory Open Access Journal |
| issn | 1475-2867 |
| language | English |
| last_indexed | 2024-12-22T19:00:17Z |
| publishDate | 2019-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer Cell International |
| spelling | doaj.art-aae3d337e416402da1cb6a4febe231ec2022-12-21T18:15:58ZengBMCCancer Cell International1475-28672019-04-0119111310.1186/s12935-019-0803-4RNF8 is responsible for ATRA resistance in variant acute promyelocytic leukemia with GTF2I/RARA fusion, and inhibition of the ubiquitin–proteasome pathway contributes to the reversion of ATRA resistanceWenzhe Yan0Ji Li1Yang Zhang2Yafei Yin3Zhao Cheng4Jiayi Wang5Guoyu Hu6Sufang Liu7Yewei Wang8Yunxiao Xu9Hongling Peng10Guangsen Zhang11Department of Hematology, The Secong Xiangya Hospital, Central South UniversityDepartment of Hematology, The Secong Xiangya Hospital, Central South UniversityDepartment of Oncology, The Secong Xiangya Hospital, Central South UniversityDepartment of Hematology, Xiangtan Central HospitalDepartment of Hematology, The Secong Xiangya Hospital, Central South UniversityDepartment of Nephrology, The Secong Xiangya Hospital, Central South UniversityDepartment of Hematology, Zhuzhou No.1 HospitalDepartment of Hematology, The Secong Xiangya Hospital, Central South UniversityDepartment of Hematology, The Secong Xiangya Hospital, Central South UniversityDepartment of Hematology, The Secong Xiangya Hospital, Central South UniversityDepartment of Hematology, The Secong Xiangya Hospital, Central South UniversityDepartment of Hematology, The Secong Xiangya Hospital, Central South UniversityAbstract Background GTF2I-RARA is a newly identified RARA fusion gene in variant acute promyelocytic leukemia (APL) patients with t(7;17)(q11;q21). Clinical manifestation in the patient showed that it is a sort of ATRA-insensitive oncogene and is different from the classic PML-RARA in terms of therapeutic reaction. Methods To reveal the functional characteristics and regulating mechanism of the GTF2I-RARA fusion gene, we established a GTF2I-RARA-transfected HL60 cell model and examined its sensitivity to ATRA by western blot, MTT assay, flow cytometry, and Wright-Giemsa staining. Coimmunoprecipitation and confocal microscopy were used to examine the binding of GTF2I-RARA and transcriptional corepressors. We also performed ChIP-seq to search for potential target genes. Immunoprecipitation, ubiquitination assay, western blot, luciferase assay, and real-time PCR were used to analyze the effects of RNF8 on RARA. Flow cytometry and Wright-Giemsa staining were used to study the effect of MG132 and ATRA on the GTF2I-RARA-transfected HL60 cell model. Result We confirmed resistance of GTF2I-RARA to ATRA. Compared with PML-RARA, GTF2I-RARA has a higher affinity to HDAC3 under ATRA treatment. Using the ChIP-sequencing approach, we identified 221 GTF2I-RARA binding sites in model cells and found that the RING finger protein 8 (RNF8) is a target gene of GTF2I-RARA. RNF8 participates in disease progression and therapy resistance in APL with the GTF2I-RARA transcript. Elevated RNF8 expression promotes the interaction between RARA and RNF8 and induces RARA Lys-48 linkage ubiquitylation and degradation, resulting in attenuated transcriptional activation of RARA. Conclusion Our results suggest that RNF8 is a key GTF2I-RARA downstream event. Using the combination of MG132 and ATRA to treat GTF2I-RARA-HL60 cells, a synergistic effect leading to GTF2I-RARA-HL60 cell differentiation was confirmed. Taken together, the targeting of RNF8 may be an alternative choice for treatment in variant APL with GTF2I-RARA fusion.http://link.springer.com/article/10.1186/s12935-019-0803-4APLGTF2I-RARARNF8RARAProteasome inhibitor |
| spellingShingle | Wenzhe Yan Ji Li Yang Zhang Yafei Yin Zhao Cheng Jiayi Wang Guoyu Hu Sufang Liu Yewei Wang Yunxiao Xu Hongling Peng Guangsen Zhang RNF8 is responsible for ATRA resistance in variant acute promyelocytic leukemia with GTF2I/RARA fusion, and inhibition of the ubiquitin–proteasome pathway contributes to the reversion of ATRA resistance Cancer Cell International APL GTF2I-RARA RNF8 RARA Proteasome inhibitor |
| title | RNF8 is responsible for ATRA resistance in variant acute promyelocytic leukemia with GTF2I/RARA fusion, and inhibition of the ubiquitin–proteasome pathway contributes to the reversion of ATRA resistance |
| title_full | RNF8 is responsible for ATRA resistance in variant acute promyelocytic leukemia with GTF2I/RARA fusion, and inhibition of the ubiquitin–proteasome pathway contributes to the reversion of ATRA resistance |
| title_fullStr | RNF8 is responsible for ATRA resistance in variant acute promyelocytic leukemia with GTF2I/RARA fusion, and inhibition of the ubiquitin–proteasome pathway contributes to the reversion of ATRA resistance |
| title_full_unstemmed | RNF8 is responsible for ATRA resistance in variant acute promyelocytic leukemia with GTF2I/RARA fusion, and inhibition of the ubiquitin–proteasome pathway contributes to the reversion of ATRA resistance |
| title_short | RNF8 is responsible for ATRA resistance in variant acute promyelocytic leukemia with GTF2I/RARA fusion, and inhibition of the ubiquitin–proteasome pathway contributes to the reversion of ATRA resistance |
| title_sort | rnf8 is responsible for atra resistance in variant acute promyelocytic leukemia with gtf2i rara fusion and inhibition of the ubiquitin proteasome pathway contributes to the reversion of atra resistance |
| topic | APL GTF2I-RARA RNF8 RARA Proteasome inhibitor |
| url | http://link.springer.com/article/10.1186/s12935-019-0803-4 |
| work_keys_str_mv | AT wenzheyan rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT jili rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT yangzhang rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT yafeiyin rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT zhaocheng rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT jiayiwang rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT guoyuhu rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT sufangliu rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT yeweiwang rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT yunxiaoxu rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT honglingpeng rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance AT guangsenzhang rnf8isresponsibleforatraresistanceinvariantacutepromyelocyticleukemiawithgtf2irarafusionandinhibitionoftheubiquitinproteasomepathwaycontributestothereversionofatraresistance |