Estradiol increases risk of topoisomerase IIβ-mediated DNA strand breaks to initiate Xp11.2 translocation renal cell carcinoma
Abstract Background Xp11.2 translocation renal cell carcinoma (tRCC) is defined by translocation of the transcription factor E3 (TFE3) gene located on chromosome Xp11.2. Due to the high incidence in women, 17β-estradiol (E2) may be a factor influencing TFE3 breaks, and the topoisomerase II (TOP2) po...
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| Format: | Article |
| Language: | English |
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BMC
2021-11-01
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| Series: | Cell Communication and Signaling |
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| Online Access: | https://doi.org/10.1186/s12964-021-00790-3 |
| _version_ | 1818826847668928512 |
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| author | Qiancheng Shi Ning Liu Lei Yang Yi Chen Yanwen Lu Hongqian Guo Xiaodong Han Dongmei Li Weidong Gan |
| author_facet | Qiancheng Shi Ning Liu Lei Yang Yi Chen Yanwen Lu Hongqian Guo Xiaodong Han Dongmei Li Weidong Gan |
| author_sort | Qiancheng Shi |
| collection | DOAJ |
| description | Abstract Background Xp11.2 translocation renal cell carcinoma (tRCC) is defined by translocation of the transcription factor E3 (TFE3) gene located on chromosome Xp11.2. Due to the high incidence in women, 17β-estradiol (E2) may be a factor influencing TFE3 breaks, and the topoisomerase II (TOP2) poison is considered one of the important risk factors in mediating DNA breaks. In this study, we investigated the potential pathogenesis of Xp11.2 tRCC using the renal epithelial cell line HK-2. Methods Immunofluorescence assay was performed to analyze DNA breaks by quantifying phosphorylation of H2AX (γH2AX), and the micronuclei (MN) assay was designed for monitoring chromosome breaks. The chromatin immunoprecipitation (CHIP) was used to detect whether proteins bound to specific DNA site, and the co-immunoprecipitation (Co-IP) was used to confirm whether proteins bound to other proteins. In some experiments, siRNA and shRNA were transfected to knockdown target genes. Results Our results demonstrated that DNA double-strand breaks were mediated by TOP2β in HK-2 cells, and this process could be amplified through estrogen receptor α (ERα)-dependent pathway induced by E2. After performing translocation site analysis using target region sequencing data in two Xp11.2 tRCC cell lines and ten Xp11.2 tRCC patients, we confirmed that TOP2β and ERα could both bind to TFE3 translocation sites directly to mediate DNA breaks in a E2-dependent manner. However, TOP2β and ERα were not observed to have direct interaction, indicating that their collaborative may be implemented in other ways. Besides, TFE3 was found to be upregulated through NRF1 with increasing E2 concentration, which could increase the risk of TFE3 breaks. Conclusion Our results indicate that E2 amplifies TOP2β-mediated TFE3 breaks by ERα-dependent pathway, and E2 upregulates TFE3 by NRF1 to increase the risk of TFE3 breaks. This suggests that E2 is an important pathogenic factor for Xp11.2 tRCC pathogenesis. Video Abstract |
| first_indexed | 2024-12-19T00:34:10Z |
| format | Article |
| id | doaj.art-fa875bb1b1e1498695558cc830fa6c45 |
| institution | Directory Open Access Journal |
| issn | 1478-811X |
| language | English |
| last_indexed | 2024-12-19T00:34:10Z |
| publishDate | 2021-11-01 |
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| series | Cell Communication and Signaling |
| spelling | doaj.art-fa875bb1b1e1498695558cc830fa6c452022-12-21T20:44:54ZengBMCCell Communication and Signaling1478-811X2021-11-0119111510.1186/s12964-021-00790-3Estradiol increases risk of topoisomerase IIβ-mediated DNA strand breaks to initiate Xp11.2 translocation renal cell carcinomaQiancheng Shi0Ning Liu1Lei Yang2Yi Chen3Yanwen Lu4Hongqian Guo5Xiaodong Han6Dongmei Li7Weidong Gan8Department of Urology, Affiliated Drum Tower Hospital, Medical School of Nanjing UniversityDepartment of Urology, Nanjing First Hospital, Nanjing Medical UniversityImmunology and Reproduction Biology Laboratory and State Key Laboratory of Analytical Chemistry for Life Science, Medical School, Nanjing UniversityImmunology and Reproduction Biology Laboratory and State Key Laboratory of Analytical Chemistry for Life Science, Medical School, Nanjing UniversityDepartment of Urology, Affiliated Drum Tower Hospital, Medical School of Nanjing UniversityDepartment of Urology, Affiliated Drum Tower Hospital, Medical School of Nanjing UniversityImmunology and Reproduction Biology Laboratory and State Key Laboratory of Analytical Chemistry for Life Science, Medical School, Nanjing UniversityImmunology and Reproduction Biology Laboratory and State Key Laboratory of Analytical Chemistry for Life Science, Medical School, Nanjing UniversityDepartment of Urology, Affiliated Drum Tower Hospital, Medical School of Nanjing UniversityAbstract Background Xp11.2 translocation renal cell carcinoma (tRCC) is defined by translocation of the transcription factor E3 (TFE3) gene located on chromosome Xp11.2. Due to the high incidence in women, 17β-estradiol (E2) may be a factor influencing TFE3 breaks, and the topoisomerase II (TOP2) poison is considered one of the important risk factors in mediating DNA breaks. In this study, we investigated the potential pathogenesis of Xp11.2 tRCC using the renal epithelial cell line HK-2. Methods Immunofluorescence assay was performed to analyze DNA breaks by quantifying phosphorylation of H2AX (γH2AX), and the micronuclei (MN) assay was designed for monitoring chromosome breaks. The chromatin immunoprecipitation (CHIP) was used to detect whether proteins bound to specific DNA site, and the co-immunoprecipitation (Co-IP) was used to confirm whether proteins bound to other proteins. In some experiments, siRNA and shRNA were transfected to knockdown target genes. Results Our results demonstrated that DNA double-strand breaks were mediated by TOP2β in HK-2 cells, and this process could be amplified through estrogen receptor α (ERα)-dependent pathway induced by E2. After performing translocation site analysis using target region sequencing data in two Xp11.2 tRCC cell lines and ten Xp11.2 tRCC patients, we confirmed that TOP2β and ERα could both bind to TFE3 translocation sites directly to mediate DNA breaks in a E2-dependent manner. However, TOP2β and ERα were not observed to have direct interaction, indicating that their collaborative may be implemented in other ways. Besides, TFE3 was found to be upregulated through NRF1 with increasing E2 concentration, which could increase the risk of TFE3 breaks. Conclusion Our results indicate that E2 amplifies TOP2β-mediated TFE3 breaks by ERα-dependent pathway, and E2 upregulates TFE3 by NRF1 to increase the risk of TFE3 breaks. This suggests that E2 is an important pathogenic factor for Xp11.2 tRCC pathogenesis. Video Abstracthttps://doi.org/10.1186/s12964-021-00790-3Renal cell carcinomaXp11.2EstradiolEstrogen receptorTopoisomeraseTFE3 |
| spellingShingle | Qiancheng Shi Ning Liu Lei Yang Yi Chen Yanwen Lu Hongqian Guo Xiaodong Han Dongmei Li Weidong Gan Estradiol increases risk of topoisomerase IIβ-mediated DNA strand breaks to initiate Xp11.2 translocation renal cell carcinoma Cell Communication and Signaling Renal cell carcinoma Xp11.2 Estradiol Estrogen receptor Topoisomerase TFE3 |
| title | Estradiol increases risk of topoisomerase IIβ-mediated DNA strand breaks to initiate Xp11.2 translocation renal cell carcinoma |
| title_full | Estradiol increases risk of topoisomerase IIβ-mediated DNA strand breaks to initiate Xp11.2 translocation renal cell carcinoma |
| title_fullStr | Estradiol increases risk of topoisomerase IIβ-mediated DNA strand breaks to initiate Xp11.2 translocation renal cell carcinoma |
| title_full_unstemmed | Estradiol increases risk of topoisomerase IIβ-mediated DNA strand breaks to initiate Xp11.2 translocation renal cell carcinoma |
| title_short | Estradiol increases risk of topoisomerase IIβ-mediated DNA strand breaks to initiate Xp11.2 translocation renal cell carcinoma |
| title_sort | estradiol increases risk of topoisomerase iiβ mediated dna strand breaks to initiate xp11 2 translocation renal cell carcinoma |
| topic | Renal cell carcinoma Xp11.2 Estradiol Estrogen receptor Topoisomerase TFE3 |
| url | https://doi.org/10.1186/s12964-021-00790-3 |
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