| 周娟,杨宝艳,张政梅.miR-18a靶向 WNT2B对子宫肌瘤细胞增殖和凋亡的影响研究[J].安徽医药,2022,26(4):795-800. |
| miR-18a靶向 WNT2B对子宫肌瘤细胞增殖和凋亡的影响研究 |
| Effect of miR-18a targeting WNT2B on the proliferation and apoptosis of uterine fibroid cells |
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| DOI：10.3969/j.issn.1009-6469.2022.04.035 |
| 中文关键词: 子宫肿瘤 平滑肌瘤 凋亡 增殖 微小核糖核酸 -18a WNT家族成员 2B |
| 英文关键词: Uterine neoplasms Leiomyoma Apoptosis Proliferation MicroRNA-18a WNT family member 2B |
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| 中文摘要: |
| 目的观察微小核糖核酸 -18a(miR-18a)靶向 WNT家族成员 2B(WNT2B)对子宫肌瘤细胞增殖和凋亡的影响。方法于 2019年 10月至 2020年 3月研究 miR-18a靶向 WNT2B对子宫肌瘤细胞增殖和凋亡的影响；子宫肌瘤组织和正常子宫肌组织收集自延安市人民医院；在子宫肌瘤细胞中转染 miR-18a模拟物( miR-18a mimics)qRT-PCR方法检测 miR-18a表达变化，四甲基偶氮唑蓝(MTT)测定细胞增殖，碘化丙啶(PI)单染法检测细胞周期变化， Annexin V-，FITC/PI双染法检测细胞凋亡变化，蛋白质印迹法( Western blotting)检测细胞周期蛋白 D1(cyclin D1)、细胞周期数依赖性蛋白激酶 4(CDK4)、B细胞淋巴瘤 /白血病 -2原癌基因(Bcl-2)、Bcl-2相关 X蛋白( Bax)蛋白表达变化。利用荧光素酶报告系统鉴定 miR-18a与 WNT2B的靶向关系。将 WNT2B过表达载体与 miR-18a mimics共转染到子宫肌瘤细胞中，同样利用上述方法测定细胞增殖、周期以及凋亡变化。结果与正常子宫肌细胞( 1.00±0.10)比较，子宫肌瘤细胞中 miR-18a表达水平( 0.52±0.06)下降；与 miR-NC组( 0.97±0.13)比较，转染 miR18a mimics后的子宫肌瘤细胞中 miR-18a表达水平( 3.21±0.25)升高，细胞增殖能力［A值( 0.44±0.06)比( 0.26±0.03)］下降，细胞 G1期比例［( 57.15±6.94)%比( 72.21±4.15)%］升高，细胞凋亡率［( 6.38±0.41)%比( 25.21±2.10)%］也升高，细胞中 cyclin D1［( 0.50±0.05)比( 0.20±0.02)］、 CDK4［( 0.54±0.06)比( 0.29±0.04)］、 Bcl-2［( 0.66±0.07)比( 0.42±0.05)］蛋白表达水平下降， Bax［(0.47±0.05)比( 0.82±0.09)］蛋白表达水平升高。 miR-18a靶向抑制子宫肌瘤细胞中 WNT2B的表达。 WNT2B过表达载体提高上调 miR-18a的子宫肌瘤细胞增殖能力，降低细胞 G1期比例，减少细胞凋亡。结论 miR-18a靶向抑制 WNT2B表达降低子宫肌瘤细胞增殖能力并诱导细胞凋亡。 |
| 英文摘要: |
| Objective To observe the effect of microRNA-18a (miR-18a) targeting WNT family member 2B (WNT2B) on the prolif eration and apoptosis of uterine fibroid cells.Methods The effect of miR-18a targeting WNT2B on the proliferation and apoptosis ofuterine fibroid cells was studied from October 2019 to March 2020. Uterine fibroids and normal uterine muscle tissues were collected from Yan'an People's Hospital. Uterine fibroid cells were transfected with miR-18a mimics, and the expression changes of miR-18a were detected by qRT-PCR. Cell proliferation was measured by methyl thiazolyl tetrazolium (MTT), cell cycle changes were detected bypropidium iodide (PI) single staining, apoptosis changes were detected by Annexin V-FITC/PI double staining, and cyclin D1, cell cy cle number-dependent protein kinase 4 (CDK4), B-cell lymphoma/leukemia-2 proto-oncogene (Bcl-2), and Bcl-2-related X protein (Bax) protein expression changes were detected by Western blotting. The targeting relationship between miR-18a and WNT2B was iden tified using the luciferase reporter system. The WNT2B overexpression vector and miR-18a mimics were cotransfected into uterine fi broid cells, and the changes in cell proliferation, cycle, and apoptosis were also measured by the above methods.Results Compared with normal uterine myocytes (1.00±0.10), the expression level of miR-18a in uterine fibroid cells (0.52±0.06) was decreased. Com pared with the miR-NC group (0.97±0.13), the expression level of miR-18a in uterine fibroid cells transfected with miR-18a mimics in creased (3.21±0.25), the cell proliferation ability [A value (0.44±0.06) vs. (0.26±0.03)] decreased, the proportion of cells in G1 phase [(57.15±6.94)% vs. (72.21±4.15)%] increased, the apoptosis rate [(6.38±0.41)% vs. (25.21±2.10)%] also increased, the protein expres sion levels of cyclin D1 [(0.50±0.05) vs. (0.20±0.02)], CDK4 [(0.54±0.06) vs. (0.29±0.04)], Bcl-2 [(0.66 ±0.07) vs. (0.42±0.05)] de creased, and the expression level of Bax [(0.47±0.05) vs. (0.82±0.09)] increased. MiR-18a inhibited WNT2B expression in uterine fi broid cells. Overexpression of WNT2B increased the proliferation of uterine fibroid cells that upregulate miR-18a, reduced the propor tion of cells in G1 phase, and decreased cell apoptosis.Conclusion The targeted inhibition of WNT2B expression by miR-18a reduces the proliferation ability of uterine fibroid cells and induces apoptosis. |
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