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In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice
Published on: August 23, 2019
MicroRNA-182 suppresses clear cell renal cell carcinoma migration and invasion by targeting IGF1R
Abstract:
The purpose of our study was aimed to determine the functional role of microRNA (miR)-182 in clear cell renal cell carcinoma (ccRCC) and try to clarify its underlying molecular mechanism. Expression of miR-182 in both cancer and peripheral blood samples was analyzed by quantitative real-time PCR (qRT-PCR). Human RCC line Caki-1 cells were transfected with miR-182 mimic, miR-182 inhibitor, or negative controls, and then the cell viability, colony-formation ability, migration, and invasion assay were determined. Luciferase reporter assay, qRT-PCR and Western blotting were used to determine whether insulin-like growth factor 1 receptor (IGF1R) was a target of miR-182. Further, small interfering RNA (siRNA) against IGF1R was co-transfected with miR-182 inhibitor into cells, and then the effects on migration and invasion were assessed. MiR-182 was down-regulated in both cancer and blood samples compared to the matched non-tumor adjacent tissues and healthy volunteers, respectively (both P<0.05). Compared to the control group, cell viability, colony-forming ability, and numbers of migrated and invaded cells were significantly decreased by transfection with miR-182 mimic but were markedly increased by miR-182 inhibitor (all P < 0.05). Luciferase reporter assay confirmed that IGF1R was a target gene of miR-182, and IGF1R was negatively regulated by miR-182. Co-transfection of miR-182 inhibitor with si-IGF1R reversed the effect of miR-182 inhibitor on the migration and invasion of the cells. MiR-182 functions as an anti-oncogene in ccRCC, and miR-182-mediated inhibition of cell migration and invasion might be through directly targeting IGF1R.
Insights
MicroRNA-182 acts as an anti-oncogene in clear cell renal cell carcinoma (ccRCC). Its downregulation promotes cancer cell migration and invasion by targeting insulin-like growth factor 1 receptor (IGF1R).
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Clear cell renal cell carcinoma (ccRCC) is a significant health concern with complex molecular underpinnings.
- MicroRNAs (miRNAs) play crucial roles in cancer development and progression, but their specific functions in ccRCC require further elucidation.
- Identifying novel therapeutic targets and understanding regulatory mechanisms are vital for advancing ccRCC treatment strategies.
Purpose of the Study:
- To investigate the functional role of microRNA (miR)-182 in clear cell renal cell carcinoma (ccRCC).
- To elucidate the molecular mechanism by which miR-182 influences ccRCC progression.
- To determine if insulin-like growth factor 1 receptor (IGF1R) is a direct target of miR-182 in ccRCC.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to analyze miR-182 expression in ccRCC tissues and blood samples.
- Cell-based assays (viability, colony formation, migration, invasion) in human RCC Caki-1 cells transfected with miR-182 mimic or inhibitor.
- Luciferase reporter assay, qRT-PCR, and Western blotting to validate IGF1R as a miR-182 target.
- Co-transfection experiments using small interfering RNA (siRNA) against IGF1R to assess its role in miR-182-mediated effects.
Main Results:
- MiR-182 was significantly downregulated in ccRCC tissues and peripheral blood compared to controls.
- Overexpression of miR-182 inhibited ccRCC cell viability, colony formation, migration, and invasion.
- Conversely, inhibition of miR-182 promoted these aggressive cellular phenotypes.
- Luciferase assays confirmed IGF1R as a direct target of miR-182, with miR-182 negatively regulating IGF1R expression.
- Silencing IGF1R expression reversed the pro-migratory and pro-invasive effects induced by miR-182 inhibition.
Conclusions:
- MiR-182 functions as a tumor suppressor (anti-oncogene) in clear cell renal cell carcinoma.
- The tumor-suppressive role of miR-182 in ccRCC is mediated, at least in part, by its direct targeting of insulin-like growth factor 1 receptor (IGF1R).
- MiR-182 inhibits ccRCC cell migration and invasion through the downregulation of IGF1R, suggesting a potential therapeutic avenue.
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