Experimental study of anti miRNA-181a induces apoptosis via Mitophagy enhancing on Multiple Myeloma cells

doi:10.3969/j.issn.1006-7795.2021.01.029

Abstract

Abstract: Objective To investigate the role and possible molecular mechanism of anti miRNA 181a in the regulation of mitophagy and apoptosis on Multiple Myeloma(MM) cells. Methods The expression of miRNA 181a in human normal peripheral blood CD138+ cells,MM patients' bone marrow CD138+ cells and human myeloma cell lines MM1S was detected by fluorescence quantitative PCR (QPCR) . MiR 181a inhibitor(miR-181a inhibitor group) and negative control group (NC group) were transfected into MM1S cells, mitochondrial membrane potential(MMP) was examined by flow cytometry, autophagy and mitochondrial structure were observed by transmission electron microscope, and Western blotting was used to detect the expression of mitophagy related protein (Cleaved-caspase-3, Bcl-2/Bax, LC3 Ⅱ/LC3-Ⅰ, P62 and parkin) in three groups. overexpression Parkin of miR-181a inhibitor group as rescue test and Western blotting was used to detect mitophagy related protein expression after overexpress Parkin. Results The relative expression levels of miR-181a in MM patients' bone marrow and MM1S cell lines were 1.57±0.09 and 1.64±0.06, which significantly higher than normal peripheral blood cell (P<0.05). After transfection of miR-181a inhibitor,the apoptosis rate of miR-181a inhibitor group is significantly higher than the other two groups, the number of autophagic vesicles decreased with mitochondrial structure badly damaged were also observed in miR-181a inhibitor group. In miR-181a inhibitor group, relative expression of autophagy related protein Cleaved-caspase-3, the ratio of Bcl-2/Bax and LC3 Ⅱ/LC3-Ⅰincreased significantly, P62 decreased remarkable (P<0.01) . After the Parkin expression was rescued, MM1S cells' viability recovered. Conclusion Anti miRNA-181a could induce apoptosis on multiple myeloma cells via enhancing the level of mitophagy.

Key words: multiple myeloma, MiRNA-181a, mitophagy, mitochondria, autophagy, apoptosis

Cui Xing, Zhong Ping. Experimental study of anti miRNA-181a induces apoptosis via Mitophagy enhancing on Multiple Myeloma cells[J]. Journal of Capital Medical University, 2021, 42(1): 167-169.

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