MicroRNA-218 inhibits gastrointestinal stromal tumor cell and invasion by targeting KIT

Abstract

The objectives of this study were to detect the expressions of microRNA-218 (miR-218) in human gastrointestinal stromal tumor (GIST) tissues and cells and explore its effects on the biological features of GIST-T1 cells and the expression of its target gene KIT, so as to provide new insights for GIST treatment. Using quantitative real-time polymerase chain reaction (qRT-PCR), we detected the expressions of miR-218 in the tissues and adjacent tissues of GIST and in the GIST cell lines including GIST882, GIST430, GIST48, and GIST-T1. Forty-eight hours after the miR-218 mimic was transfected into the GIST-T1 cells, the expression of miR-218 in the GIST-T1 cells was detected by qRT-PCR. The effect of miR-218 on the GIST-T1 cell viability was detected using MTT. The effect of miR-218 on the proliferation and apoptosis of GIST-T1 cell was analyzed using flow cytometry. Transwell invasion chamber was applied to detect the effect of miR-218 on the invasion of GIST-T1 cells. KIT was identified to be a target gene of miR-218 by the luciferase reporter enzyme system, and the effect of miR-218 on the expression of KIT protein in cells was determined using Western blotting. As shown by qRT-PCR, compared with that in the GIST adjacent tissue, the expressions of miR-218 in the tumor tissue and GIST cell lines were significantly decreased (P < 0.0001). Compared with the control group, the expression of miR-218 increased significantly in GIST-T1 cells transfected with miR-218 mimic for 48 h (P < 0.01). MTT showed that the cell viability decreased significantly after the overexpression of miR-218 in the GIST-T1 cells (P < 0.01). Flow cytometry showed that the cell proliferation index significantly declined after the overexpression of miR-218 (P < 0.01); meanwhile, the apoptosis of cells also significantly increased (P < 0.01). Detection using the Transwell invasion chamber showed that the number of cells passing through the Transwell chamber significantly dropped after the enhanced expression of miR-218 (P < 0.01). Luciferase reporter gene assay showed that, compared with the control group, the relative luciferase activity significantly declined in the miR-218 mimic transfection group (P < 0.01). Compared with the control group, the expression of KIT protein in the GIST-T1 cells transfected with miR-218 mimic for 48 h significantly decreased (P < 0.01). In conclusion, the expression of miR-218 decreases in human GIST tissue and cell lines. miR-218 can negatively regulate the expression of KIT protein and inhibit the proliferation and invasion of GIST cells. Treatment based on the enhanced expression of miR-218 may be a promising strategy for GIST.

Downregulation of miRNA-30c and miR-203a is associated with hepatitis C virus core protein-induced epithelial–mesenchymal transition in normal hepatocytes and hepatocellular carcinoma cells

“…Real-time quantitative PCR was performed using SYBR® Premix Ex Taq™ II (RAKARA, Japan) [15]. The forward primers used for miR-30c and miR-203a amplification were the sequences of mature miR-30c and miR-203a and the reverse primers were provided by the kit.…”

“…Cells were infected with 10 MOI of Ad-HCV-Core viruses or 10 MOI of Ad-blank as a control. 36 hrs later, cells were subjected to MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide) assay [15]. The optical density was read in a plate reader at 550 nm wavelength.…”

Placental Growth Factor-1 and -2 Induce Hyperplasia and Invasiveness of Primary Rheumatoid Synoviocytes

“…Real-time PCR confirmed differential expression of the following eight representative genes related to these processes (Fig. 1D): CDK1 and E2F2 involving cell proliferation (32); CASP1, CASP7, and BCL2 involving apoptosis; and NANOS1, KIT, and CCL28 involving cell migration and invasion (33)(34)(35). These data indicate that PlGF may function as a regulator of proliferation, apoptosis, and migration of RA-FLSs.…”

MiR-218 Mediates tumorigenesis and metastasis: Perspectives and implications

“…MiR-218 may also block AKT activation by targeting Rictor, independent of PI3K, PTEN or EGFR, resulting in cell growth deceleration and an increase in chemo-sensitivity of medulloblastoma [46], cervical cancer [47] and oral squamous cell carcinoma [48].…”