CircRTN4调节miR-224-5p/MYT1L轴对胶质母细胞瘤细胞恶性生物学行为的影响

doi:10.3969/j.issn.1006-6233.2024.01.03

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摘要 目的: 探讨环状RNA(CircRNA)RTN4调节微小RNA(miR)-224-5p/髓磷脂转录因子1样(MYT1L)轴对胶质母细胞瘤(GM)细胞恶性生物学行为的影响。方法: 以对数生长期的U251细胞进行设置分组:空白组、阴性对照(sh NC)组、沉默circRTN4 shRNA(sh circRTN4)组、sh circRTN4+抑制剂对照(inhibitor NC)组、sh circRTN4+miR-224-5p 抑制剂(miR-224-5p inhibitor)组。Transwell实验、流式细胞术、CCK-8、qRT-PCR、Western blot以及双荧光素酶分别检测细胞迁移与侵袭、凋亡、增殖、CircRTN4、miR-224-5p、MYT1L mRNA表达、增殖蛋白(ki-67)、凋亡蛋白(cleaved Caspase-3)、MYT1L蛋白表达以及miR-224-5p分别与CircRTN4、MYT1L靶向关系验证;qRT-PCR检测GM细胞系U118、U87、U251和LN229细胞及正常人星形胶质细胞系NHA细胞中CircRTN4、miR-224-5p、MYT1L mRNA表达水平。结果: U118、U87、U251和LN229细胞中CircRTN4、MYT1L mRNA表达较NHA细胞显著增加,miR-224-5p表达显著下降(P<0.05);miR-224-5p分别与CircRTN4、MYT1L的有靶向关系。与空白组、sh NC组相比,sh circRTN4组迁移、侵袭数、24h、48h A450值、ki-67、CircRTN4、MYT1L mRNA及蛋白表达显著降低,凋亡率、cleaved Caspase-3表达显著增加(P<0.05);与sh circRTN4+inhibitor NC组相比,sh circRTN4+miR-224-5p inhibitor组迁移、侵袭数、24h、48h A450值、ki-67、MYT1L mRNA及蛋白表达显著增加,凋亡率、cleaved Caspase-3表达显著下降(P<0.05),CircRTN4表达无统计学差异(P>0.05),体内实验证明干扰CircRTN4可显著抑制移植瘤裸鼠肿瘤质量(P<0.05)。结论: 沉默CircRTN4调节miR-224-5p/MYT1L轴抑制GM细胞恶性生物学行为。

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关键词 ：胶质母细胞瘤, 环状RTN4, miR-224-5p/MYT1L轴, 恶性生物学行为

Abstract：Objective: To investigate the effect of circRNA RTN4 (CircRNA) on the malignant biological behavior of glioblastoma (GM) cells by regulating miR-224-5p/myelin transcription factor-1-like (MYT1L) axis. Methods: U251 cells in logarithmic growth phase were divided into the following groups: blank group, negative control (sh NC) group, silenced CircRTN4 shRNA (sh CircRTN4) group, sh CircRTN4 + inhibitor control (inhibitor NC) group, and sh CircRTN4 + miR-224-5p inhibitor (miR-224-5p inhibitor) group. Transwell experiment, flow cytometry, CCK-8, qRT-PCR, Western blot, and dual-luciferase assay were used to detect cell migration and invasion, apoptosis, proliferation, CircRTN4, miR-224-5p, MYT1L mRNA expression, proliferation protein (ki-67), apoptosis protein (cleaved Caspase-3), and MYT1L protein expression. The targeting relationships between miR-224-5p and CircRTN4, MYT1L were verified. qRT-PCR was used to detect the mRNA expression levels of CircRTN4, miR-224-5p, and MYT1L in GBM cell lines (U118, U87, U251, LN229) and normal human astrocytes (NHA) cells. Results: In U118, U87, U251, and LN229 cells, CircRTN4 and MYT1L mRNA expression increased significantly, while miR-224-5p expression decreased significantly compared to NHA cells (P<0.05). miR-224-5p had targeting relationships with CircRTN4 and MYT1L. Compared with the blank group and sh NC group, the sh CircRTN4 group showed significantly decreased migration, invasion, 24h, 48h A450 values, ki-67, CircRTN4, MYT1L mRNA, and protein expression, increased apoptosis rate, and cleaved Caspase-3 expression (P<0.05). Compared with the sh CircRTN4 + inhibitor NC group, the sh CircRTN4 + miR-224-5p inhibitor group showed significantly increased migration, invasion, 24h, 48h A450 values, ki-67, MYT1L mRNA, and protein expression, decreased apoptosis rate, and cleaved Caspase-3 expression (P<0.05). CircRTN4 expression showed no statistical difference (P>0.05). In vivo experiments demonstrated that interfering with CircRTN4 significantly inhibited tumor mass in nude mice (P<0.05). Conclusion: Silencing CircRTN4 regulates the miR-224-5p/MYT1L axis and inhibits the malignant biological behavior of GM cells.

Key words： Glioblastoma CircRTN4 MiR-224-5p/MYT1L axis Malignant biological behavior

基金资助:四川省医学(青年创新)科研课题立项项目,(编号:Q21028)

通讯作者: 陈星宇

引用本文:

刘应刚, 李维民, 龙勇, 向城卫, 张施远, 陈星宇. CircRTN4调节miR-224-5p/MYT1L轴对胶质母细胞瘤细胞恶性生物学行为的影响[J]. 河北医学, 2024, 30(1): 15-22.
LIU Yinggang, LI Weimin, LONG Yong, et al. Effects of CircRTN4 on the Malignant Biological Behavior of Glioblastoma Cells by Regulating the miR-224-5p/MYT1L Axis. HeBei Med, 2024, 30(1): 15-22.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2024.01.03 或 http://www.hbyxzzs.cn/CN/Y2024/V30/I1/15

[1] Yuan F,Zhang S,Sun Q,et al.Hsa_circ_0072309 enhances autophagy and TMZ sensitivity in glioblastoma[J].CNS Neurosci Ther,2022,28(6):897-912.
[2] Liu J,Zhao K,Huang N,et al.Circular RNAs and human glioma[J].Cancer Biol Med,2019,16(1):11-23.
[3] Wong C H,Lou U K,Fung F K,et al.CircRTN4 promotes pancreatic cancer progression through a novel CircRNA-miRNA-lncRNA pathway and stabilizing epithelial-mesenchymal transition protein[J].Mol Cancer,2022,21(1):10-24.
[4] Zheng S Q,Qi Y,Wu J,et al.CircPCMTD1 acts as the sponge of miR-224-5p to promote glioma progression[J].Front Oncol,2019,9(1):398-410.
[5] Yu Z,Liu Y,Li Y,et al.miRNA-338-3p inhibits glioma cell proliferation and progression by targeting MYT1L[J].Brain Res Bull,2022,179(1):1-12.
[6] Cao Q,Shi Y,Wang X,et al.Circular METRN RNA hsa_circ_0037251 promotes glioma progression by sponging miR-1229-3p and regulating mTOR expression[J].Sci Rep,2019,9(1):19791-19800.
[7] Gao Y,Gao J,Lin F,et al.CircRNAs in tumor radioresistance[J].Biomolecules,2022,12(11):1586-1593.
[8] Qi Y,Ma N,Chen X,et al.CircRtn4 acts as the sponge of miR-24-3p to promote neurite growth by regulating CHD5[J].Front Mol Neurosci,2021,14(1):660429-660440.
[9] Li J,Sun D,Pu W,et al.Circular RNAs in cancer:biogenesis,function and clinical significance[J].Trends Cancer,2020,6(4):319-336.
[10] Li J,Liu X,Li C,et al.miR-224-5p inhibits proliferation,migration,and invasion by targeting PIK3R3/AKT3 in uveal melanoma[J].Cell Biochem,2019,120(8):12412-12421.
[11] Zhang Q,Chen S,Zhen Y,et al.Circular RNA circFGFR1 functions as an oncogene in glioblastoma cells through sponging to hsa-miR-224-5p[J].Immunol Res,2022,2022(1):251-264.
[12] Zang C S,Huang H T,Qiu J,et al.MiR-224-5p targets EGR2 to promote the development of papillary thyroid carcinoma[J].Eur Rev Med Pharmacol Sci,2020,24(9):4890-4900.
[13] Wang B,Li D,Yao Y,et al.The crucial role of DNA-dependent protein kinase and myelin transcription factor 1-like protein in the miR-141 tumor suppressor network[J].Cell Cycle,2019,18(21):2876-2892.