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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
miR-25 expression is upregulated in pancreatic ductal adenocarcinoma and promotes cell proliferation by targeting
Huimin Lu1, Ling Zhang1, Shan Lu1
1Department of Pancreatic Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, PR. China.
Abstract:
MicroRNAs (miRNAs) are reported to play a critical role in the regulation of cancer cell proliferation; however, the role of microRNA-25 (miR-25) in pancreatic ductal adenocarcinoma (PDAC) remains unclear. In the present study, the role of miR-25 in PDAC cell proliferation was investigated. Upregulated expression of miR-25 was found in PDAC tissues and cell lines by reverse transcription-quantitative PCR. Cell proliferation was significantly enhanced by overexpression of miR-25 as shown by CCK-8 assay results. Meanwhile, overexpression of miR-25 also promoted G1-to-S phase transition of the cell cycle in Aspc-1 cells via flow cytometry analysis. However downregulation of miR-25 inhibited the tumor cell proliferation and cell cycle transition. Online software was used to predict the target gene for miR-25 and luciferase reporter assay confirmed that Abl interactor 2 (ABI2) was a target of miR-25 via direct binding of its 3' untranslated region with miR-25. Moreover, results of the western blot analysis demonstrated that miR-25 negatively regulated the expression of ABI2 at the protein level. In addition, introduction of ABI2 mRNA into cells overexpressing miR-25 attenuated the carcinogenic effects of miR-25. In conclusion, these findings demonstrate that miR-25 plays an oncogenic role and promotive role in PDAC cell proliferation via targeting of ABI2.
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