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投稿时间：2020-01-21 网络发布日期：2020-10-20
投稿时间：2020-01-21 网络发布日期：2020-10-20
中文摘要: 目的 研究长链非编码RNA(lncRNA)LINC00324和微小核糖核酸(microRNA,miR)-375对胶质瘤细胞增殖、迁移、侵袭和凋亡的影响及机制。
方法 Real-time PCR检测2017年6月至2018年12月20例行手术切除患者的胶质瘤组织和脑外伤20例行颅内减压手术患者的正常脑组织中lncRNA LINC00324 和miR-375的水平,双荧光素酶报告系统验证lncRNA LINC00324与miR-375的关系。在外购的胶质瘤U251细胞中转染si-LINC00324、miR-375以抑制LINC00324和过表达miR-375;CCK8法、流式细胞术和Transwell实验分别检测胶质瘤U251细胞增殖、凋亡率、细胞迁移和侵袭能力。
结果 与正常脑组织相比,胶质瘤组织中lncRNA LINC00324表达水平显著升高,miR-375表达水平显著降低(P均<0.01)。过表达lncRNA LINC00324可下调胶质瘤U251细胞中miR-375含量(P<0.05),抑制lncRNA LINC00324可上调miR-375含量(P<0.05),即lncRNA LINC00324靶向负调控miR-375的表达。
抑制lncRNA LINC0032后，U251细胞中LINC00324含量、反映细胞增殖情况的OD值、侵袭和迁移细胞数降低，凋亡率升高（P<0.05，P<0.01）；过表达miR-375后，U251细胞中miR-375含量升高，OD值、侵袭和迁移细胞数降低，凋亡率升高（P均<0.01）；即抑制lncRNA LINC00324和过表达miR-375均可抑制U251细胞增殖、迁移、侵袭并促进细胞凋亡。在抑制LINC00324的同时干扰miR-375，U251细胞中miR-375含量降低，OD值、侵袭和迁移细胞数升高，凋亡率降低（P均<0.01），
即干扰miR-375可逆转抑制lncRNA LINC00324对U251细胞增殖、侵袭、迁移和凋亡的影响。
结论 LncRNA LINC00324通过靶向miR-375调控U251细胞的增殖、迁移、侵袭和凋亡,其可能作为胶质瘤的潜在分子靶点。
Abstract:Objective To investigate the effects of long-noncoding RNAs(lncRNA) LINC00324 and microRNA-375 (miR-375) on the proliferation,migration,invasion and apoptosis of glioma cells and its mechanism.
Methods From June 2017 to December 2018,the expression levels of lncRNA LINC00324 and miR-375 were detected by Real-time PCR respectively in glioma tissues of 20 patients with glioma resected(glioma group) and normal brain tissues from 20 patients undergoing intracranial decompression(control group),and the relationship between lncRNA LINC00324 and miR-375 was verified by the dual-luciferase reporter assay system.After Glioma U251 cells from ATCC were transfected with si-LINC00324 and miR-375 for LINC00324 inhibition and miR-375 overexpression.The proliferation,apoptosis rate,cell migration and invasion abilities of U251 cells were respectively determined by CCK8 assay,flow cytometry and Transwell assay.
Results Compared with control group,the level of lncRNA LINC00324 remarkably increased (P<0.01) and the level of miR-375 significantly decreased in glioma group (P<0.01).Overexpression of lncRNA LINC00324 could down-regulate the content of miR-375 in glioma U251 cells(P<0.05),and inhibition of lncRNA LINC00324 could up-regulate the content of miR-375 (P<0.05),that is,lncRNA LINC00324 targeting negatively regulated the expression of miR-375.After inhibiting lncRNA LINC0032,the content of LINC00324 in U251 cells,the OD value reflecting cell proliferation and the number of invading and migrating cells decreased,and the apoptosis rate increased (P<0.05,P<0.01);after overexpression of miR-375 in U251 cells,the content of miR-375 increased,the OD value,the number of invading and migrating cells decreased,and the apoptosis rate increased (all P<0.01);that is,inhibition of lncRNA LINC00324 and overexpression of miR-375 could inhibit U251 cell proliferation,migration,invasion and promote cell apoptosis.Interference with miR-375 while inhibiting LINC00324,the content of miR-375 in U251 cells decreased,the OD value,the number of invading and migrating cells increased,and the apoptosis rate decreased (all P<0.01),in other words, interference with miR-375 could reverse the effects of inhibiting lncRNA LINC00324 on the proliferation,invasion,migration and apoptosis of U251 cells.
Conclusion LncRNA LINC00324 regulates the proliferation,migration,invasion and apoptosis of U251 cells by targeting miR-375 and may be a potential molecular target for glioma.
keywords: Glioma U251 cells Long non-coding RNA LINC00324 microRNA-375 Proliferation Migration Invasion Apoptosis
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附件
| 作者 | 单位 |
| 李清泉1 | 1.南京医科大学附属逸夫医院神经外科,江苏南京210000； |
| 张雅旋1 | 1.南京医科大学附属逸夫医院神经外科,江苏南京210000； |
| 仇诚2 | 2.南京脑科医院神经外科,江苏南京210000； |
| 黄保胜1 | 1.南京医科大学附属逸夫医院神经外科,江苏南京210000 |
| Author Name | Affiliation |
| LI Qing-quan*,ZHANG Ya-xuan,QIU Cheng,HUANG Bao-sheng | *Department of Neurosurgery,Affiliated Yifu Hospital of Nanjing Medical University,Nanjing,Jiangsu 210000,China |
引用文本：
李清泉,张雅旋,仇诚,等.lncRNA LINC00324靶向miR-375对胶质瘤细胞增殖、凋亡、迁移和侵袭的影响[J].中国临床研究,2020,33(10):1302-1307.
李清泉,张雅旋,仇诚,等.lncRNA LINC00324靶向miR-375对胶质瘤细胞增殖、凋亡、迁移和侵袭的影响[J].中国临床研究,2020,33(10):1302-1307.