Abstract
BackgroundXp11.2 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.MethodsImmunofluorescence 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.ResultsOur 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.ConclusionOur 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.CY-qF8wtsVFYGeFR37VPNvVideo
Highlights
Xp11.2 translocation renal cell carcinoma is defined by translocation of the transcription factor E3 (TFE3) gene located on chromosome Xp11.2
E2 induces TFE3 breaks in HK‐2 cells Considering the potential oncogenic role of E2 in the pathogenesis of Xp11.2 translocation renal cell carcinoma (tRCC), we assessed the ability of E2 to induce DNA breaks
The renal epithelial cell line HK-2 was treated with E2 at a concentration of 10 nM, and the histone H2AX phosphorylation was used as an indicator for reflecting DNA double-strand breaks
Summary
Xp11.2 translocation renal cell carcinoma (tRCC) is defined by translocation of the transcription factor E3 (TFE3) gene located on chromosome Xp11.2. Xp11.2 tRCC is characterized by poor prognosis with local invasive and distant metastases [6,7,8,9], and the high prevalence of young adults increased the medical care related to complications of this disease in recent years. Another important feature of Xp11.2 tRCC is the female predominance. Considering age- and sex-specific factors of this type of RCC, we suspect that estrogen may play an important role in the pathogenesis of Xp11.2 tRCC
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