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中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (3): 247-253.doi: 10.12047/j.cjap.6068.2021.021

• 研究论文 • 上一篇    下一篇

miR-125b-5p对人血管瘤内皮细胞HemES增殖和凋亡的影响及其机制

王恬, 张福林, 赵越, 郭东栋, 杨瑞   

  1. 延安市人民医院肿瘤血液科, 陕西 延安 716000
  • 出版日期:2021-05-28 发布日期:2021-08-09
  • 通讯作者: Tel: 15029688651; E-mail: y0964f@163.com
  • 基金资助:
    *陕西省科技攻关项目(2016SF-138)

Effects of miR-125b-5p on the proliferation and apoptosis of human hemangioma endothelial cells HemES and its mechanism

WANG Tian, ZHANG Fu-lin, ZHAO Yue, GUO Dong-dong, YANG Rui   

  1. Department of Tumor Hematology, Yan'an People's Hospital, Yanan 716000, China
  • Online:2021-05-28 Published:2021-08-09

摘要: 目的: 研究miR-125b-5p 对人血管瘤内皮细胞HemECs增殖、凋亡的影响。方法: RT-qPCR检测人血管瘤内皮细胞HemECs及其旁系组织细胞中miR-125b-5p与MCL-1 mRNA的表达;选取HemECs细胞分为对照组、miR-NC组、miR-125b-5p mimic组、miR-125b-5p inhibitor组、pc-MCL-1组、miR-125b-5p+ pc-MCL-1组,每组设9复孔。将100 nmol · L-1 的miR-NC、miR-125b-5p mimic、miR-125b-5p inhibitor 、pc-MCL-1质粒分别或联合转染进入HemECs细胞。MTT法检测HemECs细胞增殖;流式细胞术检测HemECs细胞凋亡; 双荧光素酶报告检测靶向关系;蛋白印迹法检测Ki67、PCNA、cleaved caspase-3、Bax、Bcl-2、p-p70s6k/ p70s6k、p-AKT/AKT、p-mTOR/ mTOR蛋白相对表达水平。结果: 通过比较miR-125b-5p在血管瘤组织和细胞中的表达水平,选择下调效果比较明显的HemECs细胞系进行后续实验。与对照组相比,miR-125b-5p mimic组HemECs细胞增殖及Ki67和PCNA表达明显减少(P<0.01),细胞凋亡率及cleaved Caspase-3、Bax表达明显升高、Bcl-2表达明显降低(P<0.01),p-AKT/AKT、p-mTOR/mTOR、p-p70S6K/p70S6K表达明显下调(P<0.01);miR-125b-5p inhibitor组HemECs细胞增殖及Ki67和PCNA表达明显增加(P<0.01),细胞凋亡率及cleaved Caspase-3、Bax表达明显降低、Bcl-2表达升高(P<0.05,P< 0.01)。miR-125b-5p靶向下调MCL-1。与miR-125b-5p mimic组相比,miR-125b-5p+ pc-MCL-1组HemECs细胞增殖及Ki67和PCNA表达明显增加(P<0.01),细胞凋亡率及cleaved Caspase-3、Bax表达明显降低、Bcl-2表达明显升高(P<0.01), p-AKT/AKT、p-mTOR/mTOR、p-p70S6K/p70S6K表达明显上调(P<0.01)。结论: miR-125b-5p抑制人血管瘤内皮细胞增殖、诱导细胞凋亡,其机制可能与靶向下调MCL-1表达,抑制AKT / mTOR通路激活等有关。

关键词: miR-125b-5p, MCL-1, 血管瘤, 增殖, 凋亡

Abstract: Objective: This article mainly studies the effects of miR-125b-5p on the proliferation and apoptosis of human hemangioma endothelial cells HemECs. Methods: RT-qPCR was used to detect the expressions of miR-125b-5p and MCL-1 mRNA in HemECs and collateral cells of human hemangioma endothelial cells. HemECs were selected and divided into control group, miR-NC group, miR-125b-5p mimic group, miR-125b-5p inhibitor group, pc-MCL-1 group, miR-125b-5p+ pc-MCL-1 group, 9 multiple holes in each group. . HemECs were transfected with 100 nmol · L-1 of miR-NC, miR-125b-5p mimic, miR-125b-5p inhibitor, pc-MCL-1 plasmids separately or in combination. MTT method was used to detect the proliferation of HemECs. The apoptosis of HemECs was detected by flow cytometry. Dual luciferase report was used to to detect targeting relationship. The relative expression levels of Ki67, PCNA, cleaved caspase-3, Bax, Bcl-2, p-p70s6k/ p70s6k, p-AKT/AKT, and p-mTOR/mTOR proteins were detected by Western blot. Results: By comparing the expression levels of miR-125b-5p in hemangioma tissues and cells, HemECs cell lines with obvious down-regulation effects were selected for follow-up experiments. Compared with the control group, the proliferation of HemECs and the expressions of Ki67 and PCNA in the miR-125b-5p mimic group were decreased significantly (P<0.01). The apoptotic rate of HemECs and the expression levels of cleaved Caspase-3 and Bax were increased significantly, while the expression of Bcl-2 was decreased significantly (P<0.01). The expression levels of p-AKT/AKT, p-mTOR/mTOR and p-p70S6K/p70S6K were down-regulated significantly (P <0.01); the proliferation of HemECs and the expressions of Ki67 and PCNA in the miR-125b-5p inhibitor group were increased significantly (P <0.01); the apoptosis rate and the expressions of cleaved Caspase-3 and Bax were decreased significantly, and the expression of Bcl-2 was increased (P<0.05, P<0.01). miR-125b-5p targeted down-regulation of MCL-1. Compared with miR-125b-5p mimic group, the proliferation of HemECs and the expressions of Ki67 and PCNA in miR-125b-5p+ pc-MCL-1 group were increased significantly (P<0.01), the apoptosis rate of HemECs and the expressions of cleaved Caspase-3 and Bax were decreased significantly, while the expression of Bcl-2 was increased (P<0.01). The expressions of p-AKT/AKT, p-mTOR/mTOR, and p-p70S6K/p70S6K was also increased significantly (P<0.01). Conclusion: miR-125b-5p inhibits the proliferation of human hemangioma endothelial cells and induces apoptosis. The mechanism may be related to the targeted down-regulation of MCL-1 expression and inhibition of AKT/mTOR pathway activation.

Key words: miR-125b-5p, MCL-1, hemangioma, proliferation, apoptosis

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