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Updated: May 16, 2026
MicroRNA In situ Hybridization for Formalin Fixed Kidney Tissues
Published on: November 30, 2013
miR-21 downregulated TCF21 to inhibit KISS1 in renal cancer
Hui Zhang1, Yan Guo, Chao Shang
1Department of Urinary Surgery, Affiliated Shengjing Hospital, China Medical University, Shenyang, China.
Objective:
To investigate whether the microRNA-21 gene (miR-21) could regulate renal cancer cells invasion by downregulation of TFC21 and KISS1.
Methods:
Quantitative real-time polymerase chain reaction was applied to evaluate the expression level of miRNA-21 in renal cancer and normal renal cell samples. The regulated effects of miR-21 to TCF21 were detected by Western blot after pre/anti-miR-21 was transfected to Caki-1 cells. The luciferase activity assay was used to reveal the predicted target gene of miR-21 was direct and specific. Small interfering RNA-TCF21 was transfected to Caki-1 cells to inhibit the expression of the TCF21 gene. Next, the expression of the KISS1 gene was detected by Western blot in Caki-1 cells with TCF21 gene silencing. The expression vector, pcDNA3.1-KISS1, was transfected to Caki-1 cells to upregulate the expression of the KISS1 gene. The invasion ability of Caki-1 cells with KISS1 overexpression was analyzed using the Transwell assay.
Results:
Our study showed that miR-21 was upregulated in human renal cell carcinoma specimens compared with its expression in normal renal cell specimens. Pre-miR-21 could upregulate the expression of miR-21 and downregulate the expression of TCF21, and anti-miR-21 showed the opposite effects. siRNA-TCF21 decreased the expression of the TCF21 protein, and the expression of KISS1 was downregulated in Caki-1 cells with TCF21 gene silencing. pcDNA3.1-KISS1 transfection upregulated the expression of the KISS1 protein, and the invasion ability of Caki-1 cells with KISS1 overexpression decreased markedly.
Conclusion:
Aberrantly expressed miR-21 might regulate the TCF21-KISS1-associated renal cell carcinoma cell invasion pathway, and this miRNA signature could offer a novel potential therapeutic strategy for renal cell carcinoma.
Insights
MicroRNA-21 (miR-21) is upregulated in renal cell carcinoma and may promote cancer cell invasion. This study reveals miR-21 downregulates TCF21 and KISS1, suggesting a new therapeutic target for kidney cancer.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Renal cell carcinoma (RCC) is a significant health concern with complex molecular underpinnings.
- MicroRNAs (miRNAs) play crucial roles in cancer development and progression.
- Understanding miRNA-mediated regulatory pathways is vital for identifying novel therapeutic targets in RCC.
Purpose of the Study:
- To investigate the role of microRNA-21 (miR-21) in regulating renal cancer cell invasion.
- To determine if miR-21 influences the expression of TCF21 and KISS1.
- To explore the potential of miR-21 as a therapeutic target in renal cell carcinoma.
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) to assess miR-21 expression in RCC and normal tissues.
- Western blot analysis to evaluate protein levels of TCF21 and KISS1 following miR-21 manipulation.
- Luciferase activity assays to confirm direct targeting of genes by miR-21.
- Transwell assays to measure the invasive capacity of renal cancer cells.
Main Results:
- miR-21 expression was significantly upregulated in human renal cell carcinoma specimens compared to normal tissues.
- Overexpression of miR-21 led to downregulation of TCF21 protein, while inhibition of miR-21 had the opposite effect.
- Silencing of TCF21 resulted in decreased KISS1 expression, and conversely, KISS1 overexpression reduced renal cancer cell invasion.
- miR-21 directly targets TCF21, which in turn affects KISS1 expression and cellular invasion.
Conclusions:
- Aberrant expression of miR-21 is implicated in the regulation of the TCF21-KISS1 pathway in renal cell carcinoma.
- This miR-21-mediated pathway influences cancer cell invasion.
- The miR-21 signature presents a potential novel therapeutic strategy for renal cell carcinoma.
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