[1]卢 帅,王贤文,杨伟丽.沉默长链非编码RNA FBXO18-AS对鼻咽癌细胞增殖和侵袭的影响及机制实验研究[J].陕西医学杂志,2025,54(5):596-601.[doi:DOI:10.3969/j.issn.1000-7377.2025.05.004]
　LU Shuai,WANG Xianwen,YANG Weili.Effect and mechanism of silencing long non-coding RNA FBXO18-AS on the proliferation and invasion of nasopharyngeal carcinoma cells[J].,2025,54(5):596-601.[doi:DOI:10.3969/j.issn.1000-7377.2025.05.004]
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沉默长链非编码RNA FBXO18-AS对鼻咽癌细胞增殖和侵袭的影响及机制实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
54
期数:
2025年5期
页码:
596-601
栏目:
基础研究
出版日期:
2025-05-05

文章信息/Info

Title:
Effect and mechanism of silencing long non-coding RNA FBXO18-AS on the proliferation and invasion of nasopharyngeal carcinoma cells
作者:
卢 帅; 王贤文; 杨伟丽
(湖南中医药大学第一附属医院耳鼻咽喉头颈外科,湖南 长沙 410007)
Author(s):
LU Shuai; WANG Xianwen; YANG Weili
(Department of Otorhinolaryngology Head and Neck Surgery,the First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)
关键词:
鼻咽癌;  长链非编码RNA FBXO18-AS;  微小RNA-185-5p;  增殖;  侵袭
Keywords:
Nasopharyngeal carcinoma;  Long non-coding RNA FBXO18-AS;  miR-185-5p;  Proliferation;  Invasion
分类号:
R 36
DOI:
DOI:10.3969/j.issn.1000-7377.2025.05.004
文献标志码:
A
摘要:
目的:探讨沉默长链非编码RNA(lncRNA)FBXO18-AS对鼻咽癌细胞增殖和侵袭的影响及其分子机制。方法:采用Lnc2Cancer v2.0数据库分析FBXO18-AS在鼻咽癌及正常鼻咽部组织中的表达量。使用实时荧光定量PCR(RT-qPCR)检测FBXO18-AS在鼻咽癌细胞C666-1、CNE-2Z、HNE-1、SUNE-1和永生化鼻咽上皮细胞NP69中的表达量,选取表达量最高的细胞进行后续研究。将FBXO18-AS小干扰RNA(siRNA)重组质粒和对照质粒分别转染HNE-1细胞,分别标记为si-FBXO18-AS组和si-NC组。通过CCK-8法和Transwell实验检测HNE-1细胞增殖和侵袭能力的变化。使用Long Target软件和双荧光素酶报告基因实验分析FBXO18-AS与miR-185-5p的靶向关系。沉默FBXO18-AS表达后,采用RT-qPCR检测miR-185-5p表达水平变化,Western blot检测磷酸化表皮生长因子受体(p-EGFR)、磷酸化信号转导及转录激活因子 1(p-STAT1)、磷酸化Janus激酶(p-JAK)、磷酸化磷脂酰肌醇-3-激酶(p-PI3K)蛋白表达水平变化。结果:FBXO18-AS在鼻咽癌组织中的表达量高于正常鼻咽部组织(P<0.01)。FBXO18-AS在鼻咽癌细胞CNE-2Z、C666-1、SUNE-1、HNE-1中的表达量高于NP69细胞,且HNE-1细胞中FBXO18-AS表达量最高(均P<0.05)。si-FBXO18-AS组FBXO18-AS表达量低于si-NC组(P<0.01)。si-FBXO18-AS组HNE-1细胞增殖能力在48、72、96、120 h低于si-NC组(均P<0.05)。si-FBXO18-AS组细胞侵袭数目低于si-NC组(P<0.01)。FBXO18-AS与miR-185-5p存在结合位点。FBXO18-AS-WT+miR-185-5p共转染的荧光素酶活性低于FBXO18-AS-WT+miR-NC(P<0.01)。si-FBXO18-AS组miR-185-5p表达量高于si-NC组(P<0.01)。与si-NC组比较,si-FBXO18-AS组HNE-1细胞中EGFR信号通路蛋白p-EGFR、p-STAT1、p-JAK、p-PI3K表达水平降低(均P<0.05)。结论:FBXO18-AS在鼻咽癌组织和细胞中高表达,沉默FBXO18-AS可通过调控miR-185-5p抑制鼻咽癌细胞增殖和侵袭。
Abstract:
Objective:To investigate the effect of silencing lncRNA FBXO18-AS on the proliferation and invasion of nasopharyngeal carcinoma cells and its molecular mechanism.Methods:The expression levels of FBXO18-AS in NPC tissues and normal nasopharyngeal tissues were analyzed using the Lnc2Cancer v2.0 database.RT-qPCR was employed to measure FBXO18-AS expression in nasopharyngeal carcinoma cell lines(C666-1,CNE-2Z,HNE-1,SUNE-1)and the immortalized nasopharyngeal epithelial cell line NP69,and the cell line with the highest FBXO18-AS expression was selected for further study.FBXO18-AS small interfering RNA(siRNA)recombinant plasmids and control plasmids were transfected into HNE-1 cells,designated as the si-FBXO18-AS group and si-NC group,respectively.Changes in the cell proliferation and invasion were evaluated using the CCK-8 and Transwell assay.The targeting relationship between FBXO18-AS and miR-185-5p was analyzed using Long Target software and dual-luciferase reporter gene assays.After silencing FBXO18-AS expression,RT-qPCR was used to detect changes in miR-185-5p expression,and Western blot was performed to measure the expression levels of phosphorylated epidermal growth factor receptor(p-EGFR),phosphorylated signal transducer and activator of transcription 1(p-STAT1),phosphorylated Janus kinase(p-JAK),and phosphorylated phosphatidylinositol-3-kinase(p-PI3K).Results:FBXO18-AS expression was significantly elevated in nasopharyngeal carcinoma tissues compared to normal nasopharyngeal tissues(P<0.01).Among the nasopharyngeal carcinoma cell lines,FBXO18-AS expression was elevated in CNE-2Z,C666-1,SUNE-1 and HNE-1 cells compared to NP69 cells,with the highest level observed in HNE-1 cells(all P<0.05).FBXO18-AS expression was reduced in the si-FBXO18-AS group compared to the si-NC group(P<0.01).The proliferation ability of HNE-1 cells in the si-FBXO18-AS group was reduced at 48,72,96 and 120 hours compared to the si-NC group(all P<0.05).The number of invasive cells in the si-FBXO18-AS group was lower than in the si-NC group(P<0.01).FBXO18-AS and miR-185-5p were found to have binding sites.The luciferase activity of FBXO18-AS-WT co-transfected with miR-185-5p was lower than that of FBXO18-AS-WT co-transfected with miR-NC(P<0.01).The expression of miR-185-5p in the si-FBXO18-AS group was higher than in the si-NC group(P<0.01).Compared with the si-NC group,the expression levels of EGFR signaling pathway proteins p-EGFR,p-STAT1,p-JAK and p-PI3K were lower in the si-FBXO18-AS group(all P<0.05).Conclusion:FBXO18-AS is overexpressed in nasopharyngeal carcinoma tissues and cells.Silencing FBXO18-AS effectively inhibits the proliferation and invasion of nasopharyngeal carcinoma cells by regulating miR-185-5p.

参考文献/References:

[1] MAI H Q,CHEN Q Y,CHEN D,et al.Toripalimab plus chemotherapy for recurrent or metastatic nasopharyngeal carcinoma:The JUPITER-02 randomized clinical trial[J].JAMA,2023,330(20):1961-1970.
[2] LV J,WEI Y,YIN J H,et al.The tumor immune microenvironment of nasopharyngeal carcinoma after gemcitabine plus cisplatin treatment[J].Nat Med,2023,29(6):1424-1436.
[3] 宇汝翠,陆智慧,李金虎,等.益气养阴通络方通过lncRNA UCA1靶向调控miR-485-5p抑制糖尿病肾病大鼠肾小管上皮细胞凋亡及炎症反应作用机制[J].陕西中医,2023,44(8):1000-1004.
[4] WANG Z,WU Y,DU Z,et al.The dual functions of non-coding RNA CRNDE in different tumors[J].Mini Rev Med Chem,2023,23(6):719-733.
[5] WEI J,MENG X,WEI X,et al.Down-regulated lncRNA ROR in tumor-educated platelets as a liquid-biopsy biomarker for nasopharyngeal carcinoma[J].J Cancer Res Clin Oncol,2023,149(8):4403-4409.
[6] XU Y,CHEN L,CHEN Y,et al.Prediction of potential biomarkers in early-stage nasopharyngeal carcinoma based on platelet rna sequencing[J].Mol Biotechnol,2023,65(7):1096-1108.
[7] ZHANG Y,ZHENG W,ZHANG L,et al.LncRNA FBXO18-AS promotes gastric cancer progression by TGF-β1/Smad signaling[J].Eur J Histochem,2023,67(2):3667.
[8] FERRER J,DIMITROVA N.Transcription regulation by long non-coding RNAs:Mechanisms and disease relevance[J].Nat Rev Mol Cell Biol,2024,25(5):396-415.
[9] HE S W,LIANG Y L,ZHANG Y,et al.LINC00173 facilitates tumor progression by stimulating RAB1B-mediated PA2G4 and SDF4 secretion in nasopharyngeal carcinoma[J].Mol Oncol,2023,17(3):518-533.
[10] MONTERO J J,TROZZO R,SUGDEN M,et al.Genome-scale pan-cancer interrogation of lncRNA dependencies using CasRx[J].Nat Methods,2024,21(4):584-596.
[11] MIAO J,CHEN B,XIAO Y,et al.Long noncoding RNA LINC00173 induces radioresistance in nasopharyngeal carcinoma via inhibiting CHK2/P53 pathway[J].Cancer Gene Ther,2023,30(9):1249-1259.
[12] YANG Y,YANG Y P,YI M L,et al.Exploring the expression of SNHG1 and its effect on the PI3K-AKT axis in nasopharyngeal cancer[J].Neoplasma,2023,70(5):670-682.
[13] ZHANG W,ZHOU X,TANG Z,et al.Knockdown of lncRNA SNHG16 attenuates the proliferation and radioresistance of nasopharyngeal carcinoma cells by mediating miR-31-5p/SFN axis[J].Radiat Res,2023,199(2):124-131.
[14] YAN J,ZHOU Q.LncRNA FOXP4-AS1 silencing inhibits metastasis and epithelial-mesenchymal transition in nasopharyngeal carcinoma via miR-136-5p/MAPK1[J].Anticancer Drugs,2023,34(10):1104-1111.
[15] PENG Y,ZHANG Y,LIU Y,et al.LINC01376 promotes nasopharyngeal carcinoma tumorigenesis by competitively binding to the SP1/miR-4757/IGF1 axis[J].IUBMB Life,2023,75(9):702-716.
[16] HOU W,XU L,SU T,et al.Hypoxia induces tumor-derived exosome SNHG16 to mediate nasopharyngeal carcinoma progression through the miR-23b-5p/MCM6 pathway[J].Appl Biochem Biotechnol,2024,196(1):275-295.
[17] 王银燕,黄翠,朱艳,等.长链非编码RNA UBE2Q1-AS1对食管癌细胞增殖和迁移的调节作用及机制实验研究[J].陕西医学杂志,2024,53(9):1172-1176.
[18] LIU D,GONG H,TAO Z,et al.LncRNA IUR downregulates miR-144 to regulate PTEN in nasopharyngeal carcinoma[J].Arch Physiol Biochem,2023,129(1):116-121.
[19] ZHOU Z,CHEN Y.LncRNA SNHG1 promotes nasopharyngeal carcinoma development via targeting miR-424-5p[J].Histol Histopathol,2023,38(8):953-963.
[20] ZHOU S,CAO C,HU J.Long non-coding RNA small nucleolar RNA host gene 4 induced by transcription factor SP1 promoted the progression of nasopharyngeal carcinoma through modulating microRNA-510-5p/centromere protein F axis[J].Biochem Genet,2023,61(5):1967-1986.
[21] ASHEM M,MOHANDESI KHOSROSHAHI E,ALIAHMADY M,et al.Non-coding RNA transcripts,incredible modulators of cisplatin chemo-resistance in bladder cancer through operating a broad spectrum of cellular processes and signaling mechanism[J].Noncoding RNA Res,2024,9(2):560-582.
[22] HONG Y,LI Z,SU Y,et al.The ceRNA mechanism of lncRNA MEG3/miR-21-5p/SPRY2 in cell proliferation and apoptosis in bladder cancer[J].Crit Rev Eukaryot Gene Expr,2024,34(1):55-68.
[23] MA Y Y,GAO W,WANG H,et al.Integrated ceRNAs regulating relationship and bioinformatics analysis to study the molecular mechanisms of the inhibition of puerarin on bladder cancer cell[J].J Asian Nat Prod Res,2025,27(3):400-420.
[24] ZHAO Z Y,CAO Y,WANG H L,et al.A risk model based on lncRNA-miRNA-mRNA gene signature for predicting prognosis of patients with bladder cancer[J].Cancer Biomark,2024,39(4):277-287.
[25] ZHOU J,XU J,CHENG L,et al.Alteration of lncRNA RHPN1-AS1 predicts clinical prognosis and regulates the progression of bladder cancer via modulating miR-485-5p[J].Int J Biol Markers,2024,39(4):284-291.
[26] LU W,HUANG Z,WANG J,et al.Long non-coding RNA DANCR accelerates colorectal cancer progression via regulating the miR-185-5p/HMGA2 axis[J].J Biochem,2022,171(4):389-398.
[27] WANG Y P,HUANG Y,HOU T,et al.LncRNA XIST acts as a ceRNA sponging miR-185-5p to modulate pancreatic cancer cell proliferation via targeting CCND2[J].Transl Cancer Res,2020,9(3):1427-1438.
[28] WEN H,LIU Z,TANG J,et al.MiR-185-5p targets RAB35 gene to regulate tumor cell-derived exosomes-mediated proliferation,migration and invasion of non-small cell lung cancer cells[J].Aging(Albany NY),2021,13(17):21435-21450.
[29] MO J,GONG Z,LIU H,et al.THAP9-AS1 promotes nasopharyngeal carcinoma progression through targeted regulation of the miR-185-5p/SOX13 axis[J].Physiol Int,2024,111(1):19-34.
[30] LUO Y,YE J,DENG Y,et al.The miRNA-185-5p/STIM1 axis regulates the invasiveness of nasopharyngeal carcinoma cell lines by modulating EGFR activation-stimulated switch from E- to N-Cadherin[J].Molecules,2023,28(2):818.

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备注/Memo

备注/Memo:
[基金项目]国家自然科学基金资助项目(81973914)
更新日期/Last Update: 2025-05-05