Mechanism of disordered subcellular localization of TFE3 fusion protein in TFE3 translocation renal cell carcinoma
10.3969/j.issn.1009-8291.2023.08.015
- VernacularTitle:TFE3基因易位相关性肾癌中TFE3融合蛋白亚细胞定位失调的机制研究
- Author:
Jun PAN
1
;
Yi CHEN
2
,
3
,
4
;
Lei YANG
2
,
3
,
4
;
Ning LIU
5
;
Yanwen LU
1
;
Wenliang MA
1
;
Weidong GAN
1
;
Dongmei LI
2
,
3
,
4
Author Information
1. Department of Urology, Affiliated Drum Tower Hospital of Medical School of Nanjing University, Nanjing 210008
2. Laboratory of Immunology and Reproduction Biology, Nanjing University School of Medicine, Nanjing 210093
3. Jiangsu Key Laboratory of Molecular Medicine, Nanjing University, Nanjing 210093
4. State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing 210093
5. Department of Urology, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, China
- Publication Type:Journal Article
- Keywords:
TFE3 fusion protein;
exon;
nuclear localization;
TFE3 translocation renal cell carcinoma
- From:
Journal of Modern Urology
2023;28(8):713-719
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To investigate the effects of the loss of exon 1 of TFE3 on nuclear localization of chimeric TFE3 protein in TFE3 translocation renal cell carcinoma (TFE3 tRCC). 【Methods】 The localization of TFE3 protein in TFE3 tRCC and clear cell renal cell carcinoma (ccRCC) were detected with immunochemistry. The exon retention of TFE3 gene in TFE3 tRCC was analyzed in databases and literatures. The plasmids containing TFE3 full-length and different-length of TFE3 exons which were constructed to pCDH-MCS-EGFP-Puro were transfected into HEK293T using Lipo FiterTM. The localization of EGFP protein in HEK293T cells were detected with confocal microscopy. The localization of TFE3 protein and truncated TFE3 protein were detected with Western blotting. The mRNA expression of the downstream genes of TFE3 protein were detected with q-PCR. 【Results】 Strong nuclear signal of TFE3 protein was observed in TFE3 tRCC, whereas TFE3 protein in ccRCC was mainly localized in cytoplasm. The results of fluorescence imaging and Western blotting showed that TFE3 full-length protein was expressed both in nucleus and cytoplasm, and the expression of truncated TFE3 protein was mainly localized in nucleus. The q-PCR analysis demonstrated that the deletion of exon 1 in TFE3 gene led to a higher transcriptional level of targeted genes of TFE3 protein. 【Conclusion】 The loss of exon 1 in TFE3 played a critical role in preventing TFE3 protein from entering the nucleus. In TFE3 tRCC, the loss of exon 1 in TFE3 gene leads to the nuclear localization of TFE3 fusion protein and activation of its downstream target genes. This mechanism promises to uncover the occurrence and development of TFE3 tRCC.
