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Updated: Apr 18, 2026

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MicroRNA-21 controls hTERT via PTEN in human colorectal cancer cell proliferation
Yang Yang1, Jing-Jing Yang, Hui Tao
1Clinical College of General Hospital of Armed Police Forces, Anhui Medical University, Hefei, 230032, China.
Abstract:
Colorectal cancer is a major health problem worldwide. The aim of this study was to determine a role of microRNA-21 (miRNA-21) in colorectal cancer (CRC) and to elucidate miRNA-21 regulation of hTERT by phosphatase and tensin homologue (PTEN). Protein and mRNA samples were extracted with 30 CRC samples and one CRC cell lines. The expression of miRNA-21, hTERT, PTEN in CRC tissues and CRC cell lines was measured by real-time fluorescent quantitative PCR, Western blotting. Cell viability was detected by MTT and cell cycle assay. In this study, we show that the expression of miRNA-21 was overexpressed in CRC tissue and CRC cell lines compared with the control group. The effects of miRNA-21 were then assessed in MTT assays through in vitro transfection with a miRNA-21 mimic or inhibitor. PTEN has been identified as a target gene of miRNA-21 in CRC cell lines. Moreover, Western blot and qRT-PCR analyses revealed that miRNA-21 increased the expression of human telomerase reverse transcriptase (hTERT) via the PTEN/ERK1/2. In addition, Western blot analyses confirmed that an inverse correlation between PTEN and hTERT levels of high miRNA-21 RNA-expressing CRC tissues and cell lines. Finally, these data indicate that miRNA-21 regulates hTERT expression via the PTEN/ERK1/2 signaling pathway, therefore controlling CRC cell line growth. MiRNA-21 may serve as a novel therapeutic target in CRC.
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