miR-15b通过靶向ABCG2信号通路抑制胶质瘤干细胞迁移及侵染

发布时间：2018-06-25 04:38

本文选题：干细胞 + 微RNAs　；参考：《中国组织工程研究》2017年33期

【摘要】：背景:mi R-15b在肿瘤起始和发展过程中发挥重要作用,于是作者推测miR-15b可能参与调解胶质瘤干细胞细胞的迁移和侵染,而这目前尚无相关报道,并且其机制也不清楚。目的:探讨mi R-15b对胶质瘤干细胞迁移和侵染的影响及相关分子机制。方法:实时荧光定量PCR检测胶质瘤组织、正常成人脑组织,胶质瘤干细胞及非胶质瘤干细胞中mi R-15b的表达情况。(1)分别以ABCG2特异性si RNA、对照si RNA转染胶质瘤干细胞;(2)分别以mi R-15抑制物、mi R-15模拟物及mi R-对照分别转染胶质瘤干细胞。转染后48 h,Transwell检测细胞迁移和侵染能力,ELISA法和明胶酶谱实验检测细胞培养上清中基质金属蛋白酶2/9蛋白量及活性变化。将胶质瘤干细胞及非胶质瘤干细胞分别注入裸鼠皮下,30 d内观察成瘤情况。结果与结论:(1)与正常脑组织相比,胶质瘤中mi R-15b的表达明显较低且与胶质瘤所处阶段呈负相关性。与非胶质瘤干细胞相比,胶质瘤干细胞中mi R-15b的表达显著下调;(2)与mi R-对照组比较,mi R-15b模拟物组细胞迁移及侵染能力显著下降(P0.01),mi R-15b抑制物组细胞迁移及侵染能力显著增加(P0.01);Target Scan生物学软件预测表明ABCG2为mi R-15b潜在的靶基因;(3)与对照si RNA组比较,ABCG2 si RNA组细胞迁移及侵染能力显著下降(P0.01);(4)ELISA检测表明,上调mi R-15b基因表达显著抑制基质金属蛋白酶2/9蛋白的表达;(5)ELISA法和明胶酶谱实验检测结果表明,ABCG2 si RNA转染可显著抑制基质金属蛋白酶2/9蛋白的活性,但不影响其表达量;(6)裸鼠体内实验表明,胶质瘤干细胞具有更强的成瘤能力;(7)结果表明,mi R-15b通过靶作用于ABCG2调控胶质瘤干细胞的迁移和侵染,上调mi R-15b表达可抑制胶质瘤干细胞的迁移和侵染。
[Abstract]:Background: mi R-15b plays an important role in the initiation and development of tumor, so the authors speculate that miR-15b may be involved in the mediation of glioma stem cell migration and infection, which has not been reported and its mechanism is not clear. Objective: to investigate the effect of mi R-15b on the migration and infection of glioma stem cells and its molecular mechanism. Methods: Real-time fluorescent quantitative PCR was used to detect glioma tissue and normal adult brain tissue. The expression of mi R-15b in glioma stem cells and non-glioma stem cells. (1) glioma stem cells were transfected with ABCG2 specific si RNAs and control siRNA, respectively; (2) glioma stem cells were transfected with mi R-15 inhibitor, mi R-15 mimics, and mi R- control cells, respectively. 48 h after transfection, the ability of cell migration and infection was detected by Transwell and the changes of matrix metalloproteinase 2 / 9 protein and activity in the supernatant of cell culture were detected by Elisa and gelatin zymogram. Glioma stem cells and non glioma stem cells were injected into nude mice subcutaneously for 30 days. Results and conclusion: (1) the expression of miR-15b in gliomas was significantly lower than that in normal brain tissues and was negatively correlated with the stage of glioma. Compared with non-glioma stem cells, The expression of mi R-15b in glioma stem cells was significantly down-regulated. (2) the migration and infection ability of the cells in the Mi R-15b mimic group was significantly decreased (P0.01). (P0.01) the cell migration and invasion ability of the MIR-15b inhibitor group was significantly increased (P0.01) Target Scan biological software. It was predicted that ABCG2 was a potential target gene of mi R-15b. (3) compared with the control group, the migration and infection ability of ABCG2 siRNA group was significantly decreased (P0.01); (4) Elisa. The up-regulation of miR-15b gene expression significantly inhibited the expression of matrix metalloproteinase 2 / 9 protein. (5) the results of Elisa and gelatinase assay showed that ABCG2si RNA transfection could significantly inhibit the activity of matrix metalloproteinase 2 / 9 protein. (6) the glioma stem cells had stronger tumorigenesis ability in nude mice, (7) the results showed that the glioma stem cells could regulate the migration and infection of glioma stem cells by targeting ABCG2. Upregulating the expression of mi-15 b can inhibit the migration and infection of glioma stem cells.
【作者单位】：郑州大学附属医院(南阳医院)南阳中心医院神经内科;郑州大学附属医院(南阳医院)南阳中心医院神经外科;郑州大学附属医院(南阳医院)南阳中心医院肿瘤内科;郑州大学附属医院(南阳医院)南阳中心医院病理科;解放军沈阳军区总医院神经内科;
【基金】：国家自然科学基金青年科学基金资助项目(81401097)~~
【分类号】：R739.41

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