Regulatory effect of miR-28-5p on apoptosis and autophagy of OCI-LY7 cells by targeting BECN1
Objective To investigate the regulatory effect of miR-28-5p on apoptosis and autophagy of OCI-LY7 cells by targeting BECN1.Methods The effects of curcumin and miR-28-5p on apoptosis and autophagy of OCI-LY7 cells were determined by terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining and Western blot,and the targeting relationship between miR-28-5p and BECN1 was determined by the on-line prediction and dual-luciferase reporter assay.Results TUNEL staining and the protein levels of caspase-3 were examined.Compared with those of the control group,curcumin treatment significantly increased the percentage of apoptotic cells(P<0.05)and the protein expression of cleaved caspase-3(P<0.05).Compared with those treated with curcumin alone,cell apoptosis was significantly attenuated by knockdown of miR-28-5p(P<0.05).Curcumin treatment significantly downregulated protein levels of BECN1 and the LC3B-Ⅱ/LC3B-Ⅰ ratio and upregulated P62 in OCI-LY7 cells(P<0.05),which were significantly attenuated by knockdown of miR-28-5p(P<0.05).Transfection of NC did not influence the regulatory effect of curcumin on their protein levels.Targeting relationship between miR-28-5p and BECN1 was predicted based on online database.The dual-luciferase reporter assay revealed that co-transfection of miR-28-5p mimic and wild-type BECN1 significantly reduced luciferase activity in the wild-type BECN1 group compared with those of NC group(P<0.01),which was not influenced in the mutant-type BECN1 group.Conclusion MiR-28-5p induced apoptosis of OCI-LY7 cells by mediating curcumin.BECN1 is the target gene of miR-28-5p,and curcumin-induced overexpression of miR-28-5p directly targets BECN1 to inhibit autophagy.
miR-28-5pBECN1human lymphoma cell line OCI-LY7cell autophagycell apoptosis
万方数据
维普
