Effect of lncRNA CYTOR targeting miR-139-5p on proliferation of triple-negative breast cancer cells and its mechanism
Objective To investigate the effect of long non-coding RNA(lncRNA)cytoskeleton regulator RNA(CYTOR)targeting microRNA-139-5p(miR-139-5p)on the proliferation of triple-negative breast cancer cells and its mechanism.Methods The triple-negative breast cancer cell line HCC1806 was cultured in vitro,and the cells in the third generation,which were in the logarithmic growth phase and had good growth status,were taken for subsequent experi-ments.The targeted binding sites of lncRNA CYTOR and miR-139-5p were predicted through the StarBase database,and the targeted relationship between lncRNA CYTOR and miR-139-5p was verified through dual luciferase reporter gene experiments.HCC1806 cells were randomly divided into the blank control group,negative control group,lncRNA CYTOR silencing group,miR-139-5p overexpression group,lncRNA CYTOR silencing+miR-139-5p overexpression group,respec-tively.Cells in the negative control group,lncRNA CYTOR silencing group,miR-139-5p overexpression group,lncRNA CYTOR silencing+miR-139-5p overexpression group,and lncRNA CYTOR silencing+miR-139-5p overexpression group,were transfected with NC mimics,lncRNA CYTOR siRNA,miR-139-5p mimics,lncRNA CYTOR siRNA+miR-139-5p mimics,respectively.After 48 h of transfection,we replaced with fresh culture medium and continued to culture cells for 24 h.cells in the blank control group were cultured routinely and not transfected.We collected the aforementioned cells from each group.The expression levels of miR-139-5p and SOX8 mRNA were measured using RT-qPCR.The cell prolifer-ation ability was measured using EdU method.The expression of ATP binding cassette transporter G2(ABCG2)and sex-determining region Y-box protein 8(SOX8)was measured by Western blotting.Results Through the starBase database prediction,there was a binding site between lncRNA CYTOR and miR-139-5p.The dual luciferase reporter gene experi-ment confirmed that lncRNA CYTOR had a targeted regulatory relationship with miR-139-5p.The relative expression lev-els of miR-139-5p in the lncRNA CYTOR silencing group,miR-139-5p overexpression group,and lncRNA CYTOR silenc-ing+miR-139-5p overexpression group were all higher than those in the blank control group and negative control group(all P<0.05).The relative expression levels of SOX8 mRNA,the proportion of EdU positive cells,and the relative expres-sion levels of ABCG2 and SOX8 proteins were all lower than those in the blank control group and negative control group(all P<0.05).The relative expression level of miR-139-5p in the lncRNA CYTOR silencing+miR-139-5p overexpression group was higher than those in the lncRNA CYTOR silencing group and miR-139-5p overexpression group(both P<0.05).The relative expression level of SOX8 mRNA,the proportion of EdU positive cells,and the relative expression lev-els of ABCG2 and SOX8 proteins were all lower than those in the lncRNA CYTOR silencing group and miR-139-5p overex-pression group(all P<0.05).However,there were no statistically significant differences in the relative expression levels of miR-139-5p and SOX8 mRNA,the proportion of EdU positive cells or the relative expression levels of ABCG2 and SOX8 proteins between the blank control group and the negative control group,and between lncRNA CYTOR silencing group and miR-139-5p overexpression group(all P>0.05).Conclusion LncRNA CYTOR regulates the expression of ABCG2 and SOX8 by targeting miR-139-5p,thus promoting the proliferation of triple-negative breast cancer cells.
triple-negative breast cancerlong non-coding RNA cytoskeletal regulatory RNAmicroRNA-139-5pY-box protein 8ATP binding box transporter G2
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