[1]邱明宪,康肖,王磊.LncRNA NEAT1靶向miR-185-5p调控骨肉瘤的机制研究[J].中华老年骨科与康复电子杂志,2023,(04):233-239.[doi:10.3877/cma.j.issn.2096-0263.2023.04.006]
　Qiu Mingxian,Kang Xiao,Wang Lei..Expression of LncRNA NEAT1 in osteosarcoma cells and study of the mechanism of cellular activity via miR-185-5p[J].Chin J Geriatr Orthop Rehabil(Electronic Edition),2023,(04):233-239.[doi:10.3877/cma.j.issn.2096-0263.2023.04.006]
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LncRNA NEAT1靶向miR-185-5p调控骨肉瘤的机制研究()
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中华老年骨科与康复电子杂志[ISSN:1674-3911/CN:11-9292/R]

卷:
期数:
2023年04期
页码:
233-239
栏目:
基础研究
出版日期:
2023-08-05

文章信息/Info

Title:
Expression of LncRNA NEAT1 in osteosarcoma cells and study of the mechanism of cellular activity via miR-185-5p
作者:
邱明宪1; 康肖1; 王磊2
053000 衡水市第四人民医院骨科1，内科2
Author(s):
Qiu Mingxian1;  Kang Xiao1;  Wang Lei2.
1Department of Orthopaedics, 2Department of internal medicine, the Fourth People’s Hospital of Hengshui City, Hengshui 053000, China
关键词:
骨肉瘤;  LncRNA NEAT1;  miR-185-5p;  细胞活力
Keywords:
Osteosarcoma;  LncRNA NEAT1;  MiR-185-5p;  Cell viability
DOI:
10.3877/cma.j.issn.2096-0263.2023.04.006
文献标志码:
A
摘要:
目的 探究LncRNA NEAT1对骨肉瘤细胞的调控机制。方法 采用qRT-PCR检测正常人成骨细胞和骨肉瘤细胞中LncRNA NEAT1的差异表达，构建siRNA-NEAT1#1、siRNA-NEAT1#2、siRNA-NEAT1#3转染至骨肉瘤细胞并验证其转染效率。通过CCK-8法、流式细胞术、Transwell实验分别检测细胞活力、凋亡、迁移和侵袭能力。使用核质分离实验验证NEAT1定位。利用生物信息学预测网站预测LncRNA NEAT1的靶基因，继而采取双荧光素酶精准靶点验证实验。miR-185-5p inhibitor和siRNA-NEAT1共同转染至骨肉瘤细胞中验证LncRNA NEAT1通过海绵结合靶基因miR-185-5p对骨肉瘤细胞生物学行为进行调控。结果 LncRNA NEAT1在骨肉瘤组织及细胞中高表达，在骨肉瘤细胞中敲低LncRNA NEAT1表达抑制细胞活力、迁移、侵袭，促进细胞凋亡，LncRNA NEAT1竞争性结合miR-185-5p，同时敲低骨肉瘤细胞LncRNA NEAT1与miR-185-5p的表达部分逆转了以上细胞行为。结论 敲低miR-185-5p的表达水平可部分逆转siRNA-NEAT1对骨肉瘤细胞活力的抑制作用
Abstract:
Objective To investigate the regulatory mechanism of LncRNA NEAT1 on osteosarcoma cells. Methods qRT-PCR was used to detect the differential expression of LncRNA NEAT1 in normal human osteoblasts and osteosarcoma cells. siRNA-NEAT1#1, siRNA-NEAT1#2 and siRNA-NEAT1#3 were constructed and transfected into osteosarcoma cells and their transfection efficiency was verified. Cell viability, apoptosis, migration and invasion ability were detected by CCK-8 assay, flow cytometry and transwell assay, respectively. NEAT1 localization was verified using nucleoplasmic isolation experiments. Predicted target genes of LncRNA NEAT1 was detected by bioinformatics prediction website, followed by dual luciferase assay to verify. Co-transfection of miR-185-5p inhibitor and siRNA-NEAT1 into osteosarcoma cells was applied to verify that LncRNA NEAT1 regulated the biological behavior of osteosarcoma cells through sponge binding of the target gene miR-185-5p. Results LncRNA NEAT1 was highly expressed in osteosarcoma cells. Knockdown of LncRNA NEAT1 expression in osteosarcoma cells inhibited cell viability, migration, invasion and promoted apoptosis. LncRNA NEAT1 sponged binding to miR-185-5p, knockdown of LncRNA NEAT1 expression with miR-185-5p in osteosarcoma cells partially reversed the above cellular behaviors. Conclusion Knockdown of miR-185-5p expression level could partially reverse the inhibitory effect of siRNA-NEAT1 on osteosarcoma cell viability.

相似文献/References:

[1]王玲,王燕,冯琛,等.骨肉瘤免疫治疗的研究进展[J].中华老年骨科与康复电子杂志,2017,(06):381.[doi:10.3877/cma.j.issn.2096-0263.2017.06.013]
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备注/Memo

备注/Memo:
基金项目：衡水市科技计划项目（2020014068Z）
更新日期/Last Update: 2023-09-01