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Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
MiRNA-200a induce cell apoptosis in renal cell carcinoma by directly targeting SIRT1
Hao Fu1, Wenke Song2, Xuancai Chen1
1Department of Urology, Affiliated Nanhua Hospital, University of South China, Hengyang, 421002, Hunan, People's Republic of China.
Abstract:
Accumulating evidence indicates that microRNAs are implicated in tumor initiation and progression through negatively regulating oncogenes or tumor suppressor genes. In the present study, we report that the expression of miR-200a was significantly lower in renal cell carcinoma (RCC) specimens and RCC cell lines. Restoration of miR-200a suppressed cell growth, arrested cell cycle progression, and promoted cell apoptosis in RCC cell lines. We next used qRT-PCR array technology to identify Sirtuin 1 (SIRT1) as one of the downregulated proteins during miR-200a overexpression in 786-O cells. Following a further assay by luciferase reporter system, SIRT1 was validated as a direct target of miR-200a. Moreover, siRNA-mediated knockdown of SIRT1 could partially phenocopy the effects of miR-200a overexpression. In contrast, overexpression of truncated SIRT1 (without an endogenous 3'-UTR) could rescue the effect of miR-200a overexpression on 786-O cells, which suggested that SIRT1 3'-UTR is targeted by miR-200a specifically. These observations provide further evidence for a critical tumor-suppressive role of the miR-200a in RCC in addition to identifying a novel regulatory mechanism, which may contribute to SIRT1 upregulation in RCC.
Insights
MicroRNA miR-200a is downregulated in renal cell carcinoma (RCC). Restoring miR-200a inhibits RCC cell growth and promotes apoptosis by targeting Sirtuin 1 (SIRT1).
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- MicroRNAs (miRNAs) play crucial roles in cancer by regulating oncogenes and tumor suppressor genes.
- Dysregulation of miRNAs is frequently observed in tumor initiation and progression.
- Renal cell carcinoma (RCC) is a significant public health concern with complex molecular underpinnings.
Purpose of the Study:
- To investigate the role of miR-200a in renal cell carcinoma (RCC).
- To identify the molecular targets of miR-200a in RCC.
- To elucidate the regulatory mechanism of miR-200a in RCC pathogenesis.
Main Methods:
- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) array technology to identify target genes.
- Luciferase reporter assay to validate direct targeting.
- Cell-based assays including cell growth, cell cycle, and apoptosis analysis.
- Small interfering RNA (siRNA) and gene overexpression studies.
Main Results:
- miR-200a expression was significantly reduced in RCC specimens and cell lines.
- Overexpression of miR-200a suppressed RCC cell proliferation, induced cell cycle arrest, and promoted apoptosis.
- Sirtuin 1 (SIRT1) was identified as a direct target of miR-200a.
- Knockdown of SIRT1 partially mimicked the tumor-suppressive effects of miR-200a.
Conclusions:
- miR-200a exhibits a critical tumor-suppressive function in renal cell carcinoma.
- miR-200a directly targets and downregulates SIRT1 in RCC.
- This regulatory axis represents a novel mechanism contributing to SIRT1 upregulation in RCC and offers potential therapeutic targets.
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