miR-182靶向调控PI3K/AKT/FOXO3a信号通路对胶质瘤干细胞生物学行为的影响及其机制研究

doi:10.11904/j.issn.1002-3070.2021.02.002

实用肿瘤学杂志 ›› 2021, Vol. 35 ›› Issue (2): 103-109.doi: 10.11904/j.issn.1002-3070.2021.02.002

miR-182靶向调控PI3K/AKT/FOXO3a信号通路对胶质瘤干细胞生物学行为的影响及其机制研究

贾晓琼¹, 孙秋颖¹, 刘晓宇¹, 信涛²

1.内蒙古自治区肿瘤医院肿瘤内科(呼和浩特 010020);
2.哈尔滨医科大学附属第二医院

收稿日期:2020-11-09 修回日期:2021-02-04 出版日期:2021-04-28 发布日期:2021-04-27
通讯作者: 信涛,E-mail:xintao1234@263.net
作者简介:贾晓琼,女,(1981-),博士,副主任医师,从事胶质瘤干细胞方向的研究。
基金资助:
1.呼和浩特市科学技术协会内蒙古自治区肿瘤医院临床应用项目(编号:kjjxm201706);2.北京市CSCO临床肿瘤学研究基金(编号:Y-sy2018-057)

miR-182 targeted regulation of PI3K/AKT/FOXO3a signaling pathway,its effect on the biological behavior of glioma stem cells and mechanism

JIA Xiaoqiong¹, SUN Qiuying¹, LIU Xiaoyu¹, XIN Tao²

1. Department of Oncology,Inner Mongalia Cancer Hospital,Hohhot 010020,China;
2. The Second Affiliated Hospital of Harbin Medical University

Received:2020-11-09 Revised:2021-02-04 Online:2021-04-28 Published:2021-04-27

摘要/Abstract

摘要： 目的观察miR-182靶向调控PI3K/AKT/FOXO3a信号通路对胶质瘤干细胞(GSCs)生物学行为的影响,并研究其作用机制。方法成功从胶质瘤患者病灶组织中分离出GSCs并予以培养、传代及鉴定,分为空白对照组(生理盐水溶液)、miR-182 mimics组(细胞转染miR-182 mimics)及阴性对照组(细胞转染无意义序列),细胞转染后倒置荧光显微镜下观察miR-182 mimics的转染效率,检测各组miR-182 mRNA表达水平及增殖、迁移、侵袭、凋亡等生物学行为改变,同时检测PI3K/AKT/FOXO3a信号通路相关蛋白(PI3K、p-AKT、FOXO3a)表达水平。结果 miR-182 mimics组miR-182 mRNA表达水平明显高于空白对照组及阴性对照组(P＜0.01),空白对照组与阴性对照组比较无统计学差异(P＞0.05);miR-182 mimics组细胞转染后24、48及72 h时的细胞增殖率及迁移、侵袭能力明显高于空白对照组及阴性对照组(P＜0.05),而细胞转染后24、48及72 h时的细胞凋亡率明显低于空白对照组和阴性对照组(P＜0.01),空白对照组与阴性对照组比较无统计学差异(P＞0.05);miR-182 mimics组胶质瘤干细胞转染miR-182 mimics后PI3K、p-AKT蛋白表达水平明显高于空白对照组和阴性对照组(P＜0.01),FOXO3a蛋白表达水平明显低于空白对照组、阴性对照组(P＜0.01),而空白对照组与阴性对照组PI3K、p-AKT、FOXO3a蛋白表达水平比较无统计学差异(P＞0.05)。结论 miR-182可能通过激活活化PI3K/AKT/FOXO3a信号通路而起到增强GSCs增殖、迁移及侵袭能力和抑制细胞凋亡的作用,可能作为临床治疗胶质瘤的新靶点。

关键词: 胶质瘤, 胶质瘤干细胞, 微小核糖核酸, 磷脂酰肌醇-3-激酶, 蛋白激酶B, 人类叉头框O蛋白3a

Abstract: Objective The Objective of this study was to observe the effect of miR-182 targeted regulation of PI3K/AKT/FOXO3a signaling pathway on the biological behavior of glioma stem cells(GSCs),and its mechanism of action. Methods GSCs were successfully isolated from the lesion tissues of glioma patients,before they were cultured,passaged and identified.GSCs were divided into the blank control group(physiological saline solution),miR-182 mimics group(cells transfected with miR-182 mimics)and negative control group(cell transfected nonsense sequence).After cells transfected,the transfection efficiency of miR-182 mimics was observed under an inverted fluorescence microscope.The expression of miR-182 mRNA and biological behavior changes such as proliferation,migration,invasion and apoptosis in each group were detected.The expression of PI3K/AKT/FOXO3a signaling pathway related proteins(PI3K,p-AKT,and FOXO3a)were also detected. Results The expression of miR-182 mRNA in the miR-182 mimics group was significantly higher than that of the blank control and negative control groups(P＜0.01).There was no significant difference between the blank control and negative control groups(P＞0.05).The cell proliferation rate,migration and invasion abilities of miR-182 mimics group at 24,48 and 72 h after transfection were significantly higher than those of blank control and negative control groups(P＜0.05).The apoptosis rate was significantly lower than that of blank control and negative control groups after transfection for 24 h,48 h and 72 h(P＜0.01).There was no statistical difference between the blank control and negative control groups(P＞0.05).After transfected with miR-182 mimics,the expression of PI3K and p-AKT protein in GSCs were significantly higher than those of blank control and negative control groups(P＜0.01),and the expression of FOXO3a protein was significantly lower than that of blank control and negative control groups(P＜0.01).The expression of PI3K,p-AKT,and FOXO3a protein was not statistical difference between the blank control and negative control groups(P＞0.05). Conclusion miR-182 may enhance the proliferation,migration and invasion of GSCs and inhibit cell apoptosis by activating the PI3K/AKT/FOXO3a signaling pathway.It may be a new target for clinical treatment of glioma.

Key words: Glioma;Glioma stem cells;Microribonucleic acid;Phosphatidylinositol-3-kinase;Protein kinase B;Human forkhead box O protein 3a

中图分类号:

R739.4

贾晓琼, 孙秋颖, 刘晓宇, 信涛. miR-182靶向调控PI3K/AKT/FOXO3a信号通路对胶质瘤干细胞生物学行为的影响及其机制研究[J]. 实用肿瘤学杂志, 2021, 35(2): 103-109.

JIA Xiaoqiong, SUN Qiuying, LIU Xiaoyu, XIN Tao. miR-182 targeted regulation of PI3K/AKT/FOXO3a signaling pathway,its effect on the biological behavior of glioma stem cells and mechanism[J]. Journal of Practical Oncology, 2021, 35(2): 103-109.

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http://journal09.magtechjournal.com/Jwk3_syzlxzz/CN/Y2021/V35/I2/103

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miR-182靶向调控PI3K/AKT/FOXO3a信号通路对胶质瘤干细胞生物学行为的影响及其机制研究

miR-182 targeted regulation of PI3K/AKT/FOXO3a signaling pathway,its effect on the biological behavior of glioma stem cells and mechanism

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