Abstract:
Objective To explore the effect of miR-200a on the proliferative and invasive activities of gastric carcinoma cell line SGC7901 through 13-catenin/TCF-4 signaling.
Methods miR-200a mimics were synthesized chemically and transfected into human gastric cancer cell line SGC7901 by Lipofectamine 2000. Real-time PCR was subsequently performed to detect the transfection effect. The expression of target protein was detected by immunofluorescence assay and Western blot. pTopFlash/pFopFlash luciferase vectors were used as a measure of β-catenin/TCF complex activity. Finally, scratch test, transwell cell invasion, MTT, and flow cytometry were used to detect the migratory, invasive, and proliferative activity of the tumor cells.
Results The location of β-catenin in cells shifted from the nucleus to the cytoplasm when the miR-200a expression increased. At the same time, TCF-4 decreased in the nucleus. The transfection induced a 2.27-fold increase in the luciferase activity of TopFlash luciferase vector, but did not affect the FopFlash lucifer- ase vector. The proliferation rate, as well as migratory and invasive abilities of SGC7901, were obviously decreased when miR-200a expression was upregulated, and the miR-200a mimics led to G0/G1-phase entry.
Conclusion Over-expression of miR-200a could sup- press the activity of the β-catenin/TCF-4 pathway, consequently inhibiting the proliferation and migratory as well as invasive ability of gastric carcinoma cells.
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