| 高娟,邢海洲.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[J].安徽医药,2022,26(4):660-665. |
| 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 |
| Effect of miR-218-5p on the proliferation and apoptosis of leukemia K562 cells by targeting BCL3 |
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| DOI：10.3969/j.issn.1009-6469.2022.04.005 |
| 中文关键词: 白血病 微小 RNA-218-5p B细胞淋巴瘤因子 3 细胞增殖 凋亡 |
| 英文关键词: Leukemia miR-218-5p BCL3 Cell proliferation Apoptosis |
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| 中文摘要: |
| 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为研究对象，分别构建过表达 miR-218-5p或沉默 BCL3的 K562细胞株，噻唑蓝(MTT)法检测细胞增殖，流式细胞仪检测细胞凋亡， Western blotting检测细胞增殖抗原(Ki67)、细胞周期蛋白 D1(Cyclin D1)和活化的半胱氨酸天冬氨酸蛋白酶(Cleaved Caspase-3)的表达水平。双荧光素酶报告基因实验和 Western blotting验证 miR-218-5p和 BCL3的靶向调控关系。结果与正常人骨髓细胞相比，白血病骨髓细胞中 miR-218-5p的表达水平( 1.00±0.13)比( 0.44±0.18)显著降低， BCL3的表达水平(1.04±0.32)(4.76±1.38)(0.26±0.05)(0.78±0.13)显著升高。过表达 miR-218-5p或沉默 BCL3均可显著抑制K562细胞CyclinD1(0.69±0.0比8)比( 0.34±0.0、5)、(0.65±0.0比7)比( 0.18±0.04)和 Ki67表达( 0.68±0.07)比( 0.23±0.04)、(0.54± |
| 英文摘要: |
| 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. |
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