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Gene expression changes induced by HA-PRCC-TFE3 in HK-2 cells

ABSTRACT: Gene expression changes induced by HA-PRCC-TFE3 in HK-2 cells

PROVIDER: PRJNA1169872 | ENA |

REPOSITORIES: ENA

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HA-PRCC-TFE3 bound regions in HA-PRCC-TFE3 inducible HK-2 cells

Project description:PRCC-TFE3 is an oncogenic chimeric transcription factor derived from human TFE3-rearranged renal cell carcinoma. To analyze its genome-wide occupancy, we developed HA-tagged PRCC-TFE3-inducible HK-2 cell lines, which expresses HA-tagged TFE3 in a doxycycline-dependent manner. We determined the binding regions of HA-PRCC-TFE3 by ChIPSeq.

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Project description:PRCC-TFE3 is an oncogenic chimeric transcription factor identified in human TFE3-rearranged renal cell carcinoma. To investigate the function of PRCC-TFE3 in vivo, we generated PRCC-TFE3 knock-in (KI) mice by inserting a loxP-STOP-loxP-PRCC-TFE3 cassette into the Rosa26 locus. When PRCC-TFE3 KI mice were crossed with Cadherin 16-Cre transgenic mice (KSP-Cre), the resulting mice developed TFE3-RCC in the kidney. Our findings revealed that hypoxia-inducible factors HIF1α and HIF2α are directly upregulated by PRCC-TFE3. To clarify the roles of HIF1α and HIF2α in TFE3-RCC development, we further crossed PRCC-TFE3 KI mice with HIF1α flox and/or HIF2α flox mice.

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PRCC-TFE3 dual-fusion FISH assay: A new method for identifying PRCC-TFE3 renal cell carcinoma in paraffin-embedded tissue.

Project description:PRCC-TFE3 renal cell carcinoma (RCC) is one of the most common types of Xp11.2 translocation renal cell carcinoma (tRCC), of which the diagnosis mainly relies on reverse transcription-polymerase chain reaction (RT-PCR) or chromosomal analysis in fresh frozen samples. Herein, we developed a new dual-fusion fluorescence in situ hybridization (FISH) probe to succinctly identify PRCC-TFE3 RCC in paraffin-embedded tissue. We immunohistochemically analyzed TFE3 and cathepsin K expression in 23 cases of Xp11.2 tRCC which had been confirmed by break-apart TFE3 FISH probe. Next, the dual-fusion FISH assay was performed on these selected cases. Twenty typical cases of clear renal cell carcinoma and 20 cases of papillary renal cell carcinoma were collected as control groups. Seven cases were finally confirmed as PRCC-TFE3 RCC by FISH detection, emerging dual-fusion signals, of which 2 cases were identified as PRCC-TFE3 RCC by RT-PCR previously. All remaining cases were negative for the PRCC-TFE3 rearrangement by FISH. The TFE3 immunohistochemistry was positive in 22/23 cases and the cathepsin K was positive in 16/23 cases. All 7 PRCC-TFE3 RCCs showed positive cathepsin K immunoreactivity. Our results reveal that PRCC-TFE3 dual-fusion FISH probe is an efficient and concise technique for diagnosing PRCC-TFE3 RCC in paraffin-embedded tissue.

| S-EPMC5614571 | biostudies-literature

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