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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
Propofol Disrupts Clear Cell Renal Cell Carcinoma Tumorigenesis by Regulating circFBXW7/miR-942 Axis
Yunfeng Huang1, Yuanbo Wu1, Shengqiang Lu1
1Department of Anesthesiology, Hubei Cancer Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Background:
Propofol is a commonly used intravenous anesthetic and has been found to perform anticancer effects in many cancers. However, the effects and mechanisms of propofol in clear cell renal cell carcinoma (ccRCC) remain largely undefined.
Methods:
The expression of circular RNA FBXW7 (circFBXW7) and miR-942 was detected by qRT-PCR. Cell proliferation, apoptosis, migration, and invasion capacities were analyzed using cell counting kit-8, colony formation, flow cytometry, and transwell assays, respectively. Western blot was used to detect the expression levels of PCNA, Cleaved-caspase 3 and MMP protein. The bindings between miR-942 and circFBXW7 were verified using RNA pull-down, dual-luciferase reporter, and RIP assays. Xenograft tumor analysis was employed to detect tumorigenesis in vivo.
Results:
Propofol alleviated cell proliferation, migration, invasion, and induced apoptosis in vitro and impeded tumor growth in vivo in ccRCC. Propofol elevated the level of circFBXW7, which knockdown reversed the anticancer effects of propofol on ccRCC cell tumorigenesis. CircFBXW7 directly bound to miR-942, and suppressed ccRCC cell malignant biological behaviors via targeting miR-942. Besides that, propofol decreased miR-942 expression, and miR-942 overexpression attenuated the effects of propofol on ccRCC cells. Moreover, propofol could regulate miR-942 expression through circFBXW7.
Conclusion:
Propofol suppressed the growth, migration, and invasion of ccRCC cells by regulating circFBXW7/miR-942 axis, suggesting a potential therapeutic strategy for the intervention of human ccRCC development.
Insights
Propofol, an anesthetic, inhibits clear cell renal cell carcinoma (ccRCC) growth by regulating the circFBXW7/miR-942 pathway. This finding suggests propofol as a potential therapeutic strategy for ccRCC intervention.
Area of Science:
- Oncology
- Anesthesiology
- Molecular Biology
Background:
- Propofol exhibits anticancer properties across various cancers.
- The specific effects and mechanisms of propofol in clear cell renal cell carcinoma (ccRCC) are not well-defined.
Purpose of the Study:
- To investigate the anticancer effects of propofol in ccRCC.
- To elucidate the underlying molecular mechanisms involving circular RNA FBXW7 (circFBXW7) and miR-942.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) for circFBXW7 and miR-942 expression.
- In vitro assays (cell counting kit-8, colony formation, flow cytometry, transwell) for cell proliferation, apoptosis, migration, and invasion.
- Western blot for protein expression (PCNA, Cleaved-caspase 3, MMP).
- RNA pull-down, dual-luciferase reporter, and RIP assays for circFBXW7-miR-942 interaction.
- Xenograft tumor analysis for in vivo tumorigenesis.
Main Results:
- Propofol inhibited ccRCC cell proliferation, migration, invasion, and induced apoptosis in vitro, while impeding tumor growth in vivo.
- Propofol increased circFBXW7 expression, which was crucial for its anticancer effects; knockdown of circFBXW7 reversed these effects.
- circFBXW7 directly targeted miR-942, suppressing ccRCC cell malignancy.
- Propofol decreased miR-942 expression, and its overexpression attenuated propofol's effects, indicating propofol regulates miR-942 via circFBXW7.
Conclusions:
- Propofol suppresses ccRCC progression by modulating the circFBXW7/miR-942 axis.
- This pathway presents a potential therapeutic strategy for human ccRCC intervention.
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