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Archive > Volume 24 Issue 10 > 2017,24(10):1093-1100. DOI:10.3872/j.issn.1007-385X.2017.10.009 Prev Next
Basic Research
Regulation of miRNA-24 on proliferation and apoptosis of the gastric cancer AGS cell through targeting CARMA3 gene
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Abstract:
Objective:To explore effect of miRNA-24 on proliferation and apoptosis of the gastric cancer AGS cell and its potential mechanism. Methods: Expressions of miRNA-24 in gastric cancer AGS, MKN and HGC27 cells as well as gastric mucosal epithelium GES-1 cell were detected by qRT-PCR. The AGS cell line with over-expression of miRNA-24 was constructed. CCK-8, flow cytometry and Western blotting assays were respectively used to mea-sure proliferation vibility of the cells, cell cycle and apoptosis, and expressions of cell cycle and apoptosis-related cyclin D1, CDK2, Bcl-2, p-IκB-α/IκB-α, p-Rb/Rb proteins. Possible target gene of miRNA-24 was forecasted and verified with dual luciferase report gene assay. Results: Expressions of miRNA-24 in the gastric cells was obvious-ly lower than that in the GES-1 cell (P<0.05). Proliferation vibility of the AGS cell at 48 h in the transfected with miRNA-24 mimic (miR-24 ) group was remarkably lower than that in the control miR-NC group ([119.62 ± 12.63]% vs [147.79 ± 11.89]%, P<0.05), an arrest of cell cycle occurred, and its early and later apoptosis rates were evidently more increased than those in the miR-NC group（early apoptosis rate:[11.32+2.27]% vs [0.57 ± 0.08]%; later apopto-sis rate:([15.56 ± 2.27]% vs [0.85 ± 0.16]%, all P<0.05). In the miRNA-24 group, expression levels of cyclin D1,CDK2, Bcl2 and p-Rb proteins were all obviously lower than those in the miR-NC group, and expression level of p-IκB-α protein was obviously higher than that in the miR-NC group. In co-transfection of miRNA-24 mimic and over-expression plasmid of CARMA3 that may be action target of miRNA-24 (miR-24+pcDNA-CARMA3) group,CARMA3 protein expression of the AGS cell was significantly higher than that in the miR-24 group (1.74 ± 0.09 vs 1.03 ± 0.06, P<0.05). Proliferation vibility of the AGS cell at 48 h in the miR-24+pcDNA-CARMA3 group was obvi-ously higher than that in the miR-24 group ([137.85 ± 15.34]% vs [102.31 ± 11.23]%, P<0.05), however, early and lat-er apoptosis rates of the AGS cell in the miR-24+pcDNA-CARMA3 group were all lower than those in the miR-24 group (early apoptosis rate: [4.24 ± 0.56]% vs [11.32 ± 2.27]%, later apoptosis rate: [6.38+0.63]% vs [15.56 ± 2.27]%,all P<0.05). Effect of miRNA-24 on proliferation of apoptosis of the gastric cancer AGS cell can be partially re-versed by over-expression of CARMA3. Conclusion: miRNA-24 could inhibit proliferation of the gastric caner cells and promote their apoptosis through targeting CARMA3.
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