miR-218-5p靶向B细胞淋巴瘤因子3表达影响白血病K562细胞的增殖和凋亡

目的 探讨微小RNA-218-5p(miR-218-5p)靶向B细胞淋巴瘤因子3(BCL3)基因对白血病细胞增殖和凋亡的影响.方法 反转录及荧光定量PCR(RT-qPCR)和蛋白质印迹法(Western blotting)检测正常人骨髓细胞和白血病骨髓细胞中miR-218-5p和BCL3的表达水平.以白血病细胞K562...

Effect of miR-218-5p on the proliferation and apoptosis of leukemia K562 cells by targeting BCL3

Objective To investigate the effect of micro-218-5p (miR-218-5p) on the proliferation and apoptosis of leukemia cells by targeting B-cell lymphoma factor 3 (BCL3) gene.Methods The expression levels of miR-218-5p and BCL3 in normal human bone marrow cells and leukemia bone marrow cells were detected by qRT-PCR and Western blot. The K562 cell lines over-expressing miR218-5p or silencing BCL3 were constructed. Cell proliferation was detected by MTT assay, apoptosis was detected by flow cytometry,the expression levels of Ki67, Cyclin D1 and Cleaved Caspase-3 were detected by Western blot. The dual luciferase reporter gene assayand Western blot were used to verify the targeted and regulatory relationship between miR-218-5p and BCL3.Results Compared with normal human bone marrow cells, the expression level of miR-218-5p (1.00±0.13) vs. (0.44±0.18) in leukemia bone marrow cells was significantly decreased, while the expression level of BCL3 (1.04±0.32) vs. (4.76±1.38), (0.26±0.05) vs. (0.78±0.13) was significantly in creased. Over-expression of miR-218-5p or silencing BCL3 significantly inhibited the expression level of Cyclin D1 (0.69±0.08 vs. 0.34±0.05), (0.65±0.07 vs. 0.18±0.04) and Ki67 (0.68±0.07) vs. (0.23±0.04), (0.54±0.06) vs. (0.24±0.04) in K562 cells, promoted the expression of Cleaved-Caspase-3(0.38±0.04) vs. (0.89±0.11),(0.17±0.04) vs (0.56±0.07), inhibited cell proliferation(98.72±12.38)% vs. (69.79±7.62)%], (97.48±11.49)% vs. (65.73±6.86)%] and promoted apoptosis(7.26±0.85)% vs. (24.46±2.67)%], (6.63±0.85)% vs. (26.46±2.81)%]. BCL3 was a target gene of miR-218-5p, and miR-218-5p negatively regulated the expression of BCL3. The over-ex pression of BCL3 partially reversed the effect of miR-218-5p on proliferation and apoptosis of K562 cells.Conclusion miR-218-5p in hibits leukemia cell proliferation and promotes apoptosis by targeting BCL3.