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Comparing Metastatic Clear Cell Renal Cell Carcinoma Model Established in Mouse Kidney and on Chicken Chorioallantoic Membrane
Published on: February 8, 2020
LINC02532 Contributes to Radiosensitivity in Clear Cell Renal Cell Carcinoma through the miR-654-5p/YY1 Axis
Xiaoguang Zhou1, Bowen Zeng1,2, Yansheng Li1
1Department of Urology, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China.
Background:
Studies have shown that long non-coding RNAs (lncRNAs) play essential roles in tumor progression and can affect the response to radiotherapy, including in clear cell renal cell carcinoma (ccRCC). LINC02532 has been found to be upregulated in ccRCC. However, not much is known about this lncRNA. Hence, this study aimed to investigate the role of LINC02532 in ccRCC, especially in terms of radioresistance.
Methods:
Quantitative real-time PCR was used to detect the expression of LINC02532, miR-654-5p, and YY1 in ccRCC cells. Protein levels of YY1, cleaved PARP, and cleaved-Caspase-3 were detected by Western blotting. Cell survival fractions, viability, and apoptosis were determined by clonogenic survival assays, CCK-8 assays, and flow cytometry, respectively. The interplay among LINC02532, miR-654-5p, and YY1 was detected by chromatin immunoprecipitation and dual-luciferase reporter assays. In addition, in vivo xenograft models were established to investigate the effect of LINC02532 on ccRCC radioresistance in 10 nude mice.
Results:
LINC02532 was highly expressed in ccRCC cells and was upregulated in the cells after irradiation. Moreover, LINC02532 knockdown enhanced cell radiosensitivity both in vitro and in vivo. Furthermore, YY1 activated LINC02532 in ccRCC cells, and LINC02532 acted as a competing endogenous RNA that sponged miR-654-5p to regulate YY1 expression. Rescue experiments indicated that miR-654-5p overexpression or YY1 inhibition recovered ccRCC cell functions that had been previously impaired by LINC02532 overexpression.
Conclusions:
Our results revealed a positive feedback loop of LINC02532/miR-654-5p/YY1 in regulating the radiosensitivity of ccRCC, suggesting that LINC02532 might be a potential target for ccRCC radiotherapy. This study could serve as a foundation for further research on the role of LINC02532 in ccRCC and other cancers.
Insights
This study reveals a LINC02532/miR-654-5p/YY1 feedback loop enhances clear cell renal cell carcinoma (ccRCC) radioresistance. Targeting LINC02532 could improve ccRCC radiotherapy outcomes.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Long non-coding RNAs (lncRNAs) are implicated in tumor progression and radiotherapy response in clear cell renal cell carcinoma (ccRCC).
- LINC02532 is upregulated in ccRCC, but its specific role, particularly in radioresistance, remains largely uncharacterized.
Purpose of the Study:
- To investigate the functional role of LINC02532 in ccRCC radioresistance.
- To elucidate the molecular mechanism underlying LINC02532's regulation of ccRCC radiosensitivity.
Main Methods:
- Quantitative real-time PCR and Western blotting to assess expression levels of LINC02532, miR-654-5p, and YY1.
- Clonogenic survival, CCK-8, and flow cytometry assays to evaluate cell radiosensitivity, viability, and apoptosis.
- Chromatin immunoprecipitation, dual-luciferase reporter assays, and in vivo xenograft models to determine the regulatory network and in vivo effects.
Main Results:
- LINC02532 expression was high in ccRCC cells and increased post-irradiation, correlating with radioresistance.
- LINC02532 knockdown significantly enhanced ccRCC cell radiosensitivity both in vitro and in vivo.
- A positive feedback loop involving YY1 activating LINC02532, and LINC02532 sponging miR-654-5p to regulate YY1, was identified.
Conclusions:
- A novel LINC02532/miR-654-5p/YY1 positive feedback loop critically regulates ccRCC radioresistance.
- LINC02532 emerges as a potential therapeutic target to enhance the efficacy of radiotherapy for ccRCC.
- This study provides a foundation for further research into LINC02532's role in ccRCC and other cancers.
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