Abstract
Purpose: To investigate the effect of dexmedetomidine (DEX) on epithelial mesenchymal transition (EMT) in gastric cancer cells, and the role of microRNA-144-3p (miR-144-3p) in the process.Methods: The effect of DEX on miRNA expression profile was analyzed using GEO database(https://www.ncbi.nlm.nih.gov/gds/). Human gastric cancer cells were cultured in vitro, and one group of cells was treated with saline for 48 h (control group). Cells treated with DEX at doses of 0.01, 0.1 and 1.0 μmol/L for 48 h were marked as low-, medium- and high-DEX concentration groups. The mRNA expression levels of miR-144-3p, ZEB1, E-cadherin and vimentin were determined using real-time quantitative polymerase chain reaction (RT-PCR), while the protein expressions of ZEB1, E-cadherin and vimentin were assayed with Western blotting. Cell proliferation was determined with CCK-8 assay, while metastasis was measured using Transwell assay.Results: The GEO database demonstrated that the expression of miR-144-3p in rat cardiomyocytes was significantly decreased after DEX treatment (p < 0.05). The expression of miR-144-3p was decreased in all groups, when compared to the control group, but the expressions of ZEB1 and vimentin were increased, while that of E-cadherin was down-regulated (p < 0.05). Cell proliferation in the high-DEX concentration group was decreased (p < 0.05). The degrees of cell invasion and migration were increased in the medium- and high-DEX concentration groups (p < 0.05).Conclusion: DEX promotes the metastasis of gastric cancer cells by regulation of epithelialmesenchymal transition (EMT) and the expression of miR-144-3p. This finding provides a new insight into the treatment of gastric cancer.
Highlights
Gastric cancer is second only to lung cancer as a leading cause of death in the world [1]
Bioinformatics methods and in vitro experiments were used to investigate the regulatory effect of DEX on gastric cancer cellrelated miRNA and malignant behavior, in order to provide a new insight into the use of DEX in malignant tumors
Effect of DEX on the proliferation of HGC-27 cells Compared with the control group, the proliferative potential of HGC-27 cells in high DEX concentration group was decreased significantly at 48 h and 72 h, while there were no significant changes in cell proliferation at the low and middle DEX concentration groups (p > 0.05; Figure 3)
Summary
Gastric cancer is second only to lung cancer as a leading cause of death in the world [1]. Antibodies for ZEB1, E-cadherin, Vimentin, β-actin were got from Proteintech Company (Proteintech, Wuhan, China), while Cell Counting Kit (CCK-8) was obtained from Topscience Co. Ltd (Shanghai, China). Matrigel glue (50 mg/L) was diluted in serum-free RPMI-1640 medium at a volume ratio of 8:1 It was evenly spread in the center of Transwell chamber, and air-dried at 37 °C for 2 h as basement membrane reservoir. Cells in each group were digested, centrifuged and resuspended in RPMI-1640 containing 1 % FBS in serum-free RPMI-1640 hydrated basement membrane. After addition of 600 μL of RPMI-1640 containing 10 % FBS solution to the lower chamber, the cells were cultured in a constant temperature incubator for 24 h.
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