| 曾胜,陈佳汝,张荣,等.微小 RNA-216b-5p对骨肉瘤 MG63细胞功能的影响及其与高迁移率族蛋白 B1的靶向关系[J].安徽医药,2021,25(8):1605-1609. |
| 微小 RNA-216b-5p对骨肉瘤 MG63细胞功能的影响及其与高迁移率族蛋白 B1的靶向关系 |
| Effect of miR-216b-5p on the function of osteosarcoma MG63 cells and its targeting relation?ship with HMGB1 |
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| DOI：10.3969/j.issn.1009-6469.2021.08.031 |
| 中文关键词: 骨肉瘤 微小 RNA-216b-5p 细胞增殖 细胞侵袭 细胞凋亡 高迁移率族蛋白 B1 |
| 英文关键词: Osteosarcoma miR-216b-5p Cell proliferation Invasion Apoptosis HMGB1 |
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| 中文摘要: |
| 目的探讨微小 RNA-216b-5p(miR-216b-5p)对骨肉瘤 MG63细胞增殖、侵袭和凋亡的影响及其与高迁移率族蛋白 B1(HMGB1)的靶向关系。方法将体外培养的 MG63细胞分为 mimics-NC组、 miR-216b-5p mimics组、 inhibitor-NC组和 miR216b-5p inhibitor组，采用实时荧光定量 PCR检测 miR-216b-5p的表达， MTT法、 Transwell小室和流式细胞仪分别检测细胞增殖、侵袭和凋亡能力。采用双荧光素酶报告基因实验检测 miR-216b-5p和 HMGB1的靶向关系，蛋白质印迹法( Western blot‐ting)检测 HMGB1蛋白的表达。结果与 mimics-NC组相比， miR-216b-5p mimics组细胞中 miR-216b-5p的表达水平［( 5.36±0.54)比( 1.00±0.11)］和细胞凋亡率［( 20.36±3.15)%比( 8.58±1.22)%］明显升高，而细胞增殖活力、侵袭能力和细胞中 HMGB1蛋白的表达水平均明显降低( P<0.05)； miR-216b-5p inhibitor组细胞中 miR-216b-5p的表达水平［(0.24±0.03)比( 0.96±0.08)］和细胞凋亡率［(2.05±0.38)比( 9.27±1.16)］较 inhibitor-NC组明显降低，而细胞增殖活力、侵袭能力和 HMGB1蛋白的表达水平较 inhibitor-NC组均明显升高( P<0.05)。双荧光素酶报告基因实验证实 HMGB1是 miR-216b-5p的靶基因。结论 miR-216b-5p可能通过靶向调控 HMGB1表达，抑制骨肉瘤 MG63细胞增殖、侵袭并诱导细胞凋亡。 |
| 英文摘要: |
| Objective To investigate the effect of miR-216b-5p on proliferation, invasion and apoptosis of osteosarcoma MG63 cells and its targeting relationship with HMGB1.Methods The MG63 cells cultured in vitro were divided into mimics-NC group, miR-216b5p mimics group, inhibitor-NC group and miR-216b-5p inhibitor group. The expression of miR-216b-5p was detected by real-time fluorescent quantitative PCR. The cell proliferation activity, cell invasion ability and apoptosis ability were measured by MTT, Transwellchamber and flow cytometry separately. The targeting relationship between miR-216b-5p and HMGB1 was detected by the dual luciferase reporter gene assay, and the expression of HMGB1 protein was detected by Western blot.Results Compared with the mimics-NC group, the expression level [(5.36±0.54) vs. (1.00±0.11)] of miR-216b-5p and apoptotic rate [(20.36±3.15)% vs. (8.58±1.22)%] were significantly increased in the miR-216b-5p mimics group, while the cell proliferation activity, invasion ability and expression level ofHMGB1 protein in the cells were significantly decreased (P<0.05); the expression level[(0.24±0.03) vs. (0.96±0.08)] of miR-216b-5p and apoptotic rate[(2.05±0.38) vs. (9.27±1.16)] in miR-216b-5p inhibitor group were significantly lower than that in inhibitor-NC group,and the cell proliferation activity, invasion ability and expression level of HMGB1 protein were significantly higher than that of inhibitor-NC group (P<0.05). The dual luciferase reporter gene assay confirmed that HMGB1 was the target gene of miR-216b-5p.Conclu? sion miR-216b-5p can inhibit the proliferation and invasion of osteosarcoma MG63 cells, and its mechanism may be related to the targeted regulation of HMGB1 expression. |
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