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通讯作者:

王迎伟,E-mail:wangyw1508@njmu.edu.cn;

赵晨卉,candy9023@163.com

中图分类号:R734.2

文献标识码:A

文章编号:1007-4368(2023)03-304-07

DOI:10.7655/NYDXBNS20230302

参考文献 1
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参考文献 2
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参考文献 3
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参考文献 4
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参考文献 5
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参考文献 6
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参考文献 7
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参考文献 8
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参考文献 9
CHENG G,LI Y,LIU Z,et al.LncRNA PSMA3⁃AS1 pro⁃ motes the progression of non ⁃ small cell lung cancer through targeting miR ⁃ 17 ⁃ 5p/PD ⁃ L1[J].Adv Clin Exp Med,2021,30(10):1043-1050
参考文献 10
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参考文献 11
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参考文献 12
ZHANG D H,ZHANG H F,WANG X L,et al.LINC01518 knockdown inhibits tumorigenicity by suppression of PIK3CA/Akt pathway in oesophageal squamous cell carci⁃ noma[J].Artif Cells Nanomed Biotechnol,2019,47(1):4284-4292
参考文献 13
KONG N,BAO Y,ZHAO H,et al.Long noncoding RNA LINC01518 modulates proliferation and migration in TGF⁃ β1 ⁃treated human tenon capsule fibroblast cells through the regulation of hsa ⁃miR ⁃ 216b ⁃5p[J].Neuromolecular Med,2022,24(2):88-96
参考文献 14
YOJIRO K,NATSUMI M,TAKAHIRO W,et al.OIP5 ⁃ AS1 promotes proliferation of non⁃small⁃cell lung cancer and head and neck squamous cell carcinoma cells[J].Cancer Genom Proteom,2021,18(4):543-548
参考文献 15
PENG X,WEI F,HU X.Long noncoding RNA DLGAP1⁃ AS1 promotes cell proliferation in hepatocellular carcino⁃ ma via sequestering miR ⁃ 486 ⁃ 5p[J].J Cell Biochem,2020,121(2):1953-1962
参考文献 16
CHEN W H,WANG L F,LI X Y,et al.LncRNA SNHG17 regulates cell proliferation and invasion by targeting miR⁃ 338⁃3p/SOX4 axis in esophageal squamous cell carcinoma [J].Cell Death Dis,2021,12(9):806
参考文献 17
WANG J,YU Z Y,WANG J,et al.LncRNA NUTM2A ⁃ AS1 positively modulates TET1 and HIF ⁃ 1A to enhance gastric cancer tumorigenesis and drug resistance by spong⁃ ing miR⁃376a[J].Cancer Med,2020,9(24):9499-9510
参考文献 18
YANG Y,YU Q,LI B,et al.BBOX1⁃AS1 accelerates gas⁃ tric cancer proliferation by sponging miR⁃3940⁃3p to up⁃ regulate BIRC5 expression[J].Dig Dis Sci,2021,66(4):1054-1062
目录contents

    摘要

    目的:探讨长链非编码RNA01518(long intergenic non-protein coding RNA 01518,LINC01518)基因过表达或沉默对白介素(interleukin,IL)-17 诱导非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞增殖的影响。方法:Western blot 检测 NSCLC细胞系(PC9、H1299和H1975)IL-17受体A(IL-17 receptor A,IL-17RA)的表达后,用IL-17刺激H1299和PC9细胞不同时间,利用CCK-8实验测定细胞的增殖水平。LncRNA芯片筛查IL-17刺激H1299和PC9细胞3 h后LncRNA上调的结果,从中选取某些上调的LncRNA进行RT-PCR和Real-time PCR验证。此外,将构建的pcDNA3.1/LINC01518或shLINC01518质粒转染 H1299细胞,用CCK-8、EdU和克隆形成实验检查细胞的增殖情况。结果:3种NSCLC细胞系均有IL-17RA的表达。实验发现, IL-17刺激H1299和PC9细胞后能提高其增殖水平,同时受刺激的细胞内LINC01518的上调最为显著,且过表达LINC01518可提高H1299细胞的增殖水平,而沉默LINC01518基因则能抑制IL-17诱导的细胞增殖。结论:LINC01518能促进IL-17诱导的 H1299细胞增殖。

    Abstract

    Objective:This study aims to explore the effect of overexpressing or silencing long intergenic non - protein coding RNA 01518(LINC01518)on the cell proliferation induced by IL -17 in non - small cell lung cancer(NSCLC). Methods:The IL -17 receptor A(IL-17RA)expression in NSCLC cell lines(PC9,H1299 and H1975)was first examined using Western blot. Then H1299 and PC9 cells were exposed to IL-17 for different time,and the cell proliferation was detected by CCK-8 assay. Based on the results of up-regulated lncRNAs detected by lncRNA microarray in IL-17-stimulated H1299 and PC9 cells,several lncRNAs were selected and verified by RT - PCR and Real - time PCR. Additionally,the constructed plasmids of pcDNA3.1/LINC01518 or shLINC01518 were transfected into H1299 cells,and H1299 cell proliferation were examined by CCK-8,EdU and colony formation assays. Results: IL-17RA could express in 3 kinds of NSCLC cell lines. The cell proliferation and LINC01518 were obviously up-regulated in H1299 and PC9 cells treated by IL-17. Besides,LINC01518 overexpression could markedly promote the cell proliferation,while LINC01518 gene knockdown followed by IL - 17 treatment did not enhance cell proliferation. Conclusion:LINC01518 can promote H1299 cell proliferation induced by IL-17 stimulation.

    关键词

    NSCLCIL-17LINC01518细胞增殖

    Keywords

    NSCLCIL-17LINC01518cell proliferation

  • 非小细胞肺癌(non ⁃ small cell lung cancer, NSCLC)是发病率较高的恶性肿瘤[1]。研究表明, NSCLC的致病因素除了遗传、环境和吸烟外,还与患者肺部的慢性炎症及促炎因子增多密切相关[2-3]。白介素(interleukin,IL)⁃17 是一种促炎细胞因子,在多种癌组织内显著表达,能促进癌细胞的异常增殖[4-6]。本课题组过去已证实,NSCLC 患者的癌组织中,IL ⁃17 及 IL ⁃17 受体 A(IL ⁃17 receptor A,IL ⁃ 17RA)均明显表达,且用IL⁃17刺激NSCLC细胞,上调的HMGA1(high mobility group protein A1)还能通过增强cyclin D1引起细胞的增殖反应[6]

  • 当细胞受到刺激后,除了信号分子、转录因子明显激活或表达外,还能测到某些长链非编码RNA (long non⁃coding RNA,LncRNA)的表达上调与下调[7-18]。LncRNA是长度≥200个核苷酸且多无编码蛋白功能的 RNA,能对基因的表达进行调控[7-18]。文献报道,LncRNA 在NSCLC 癌组织内表达可影响癌细胞的侵袭、转移及增殖等多个过程[9-10]

  • 长链非编码RNA 01518(long intergenic non⁃pro⁃ tein coding RNA 01518,LINC01518)是一种基因间的LncRNA,能参与不同疾病的发生与发展。例如:在食管癌组织中,LINC01518的表达上调能竞争结合miR⁃1⁃3p,诱导细胞的过量增殖[11-12]。此外,在青光眼患者肌腱囊成纤维细胞中,LINC01518也可海绵化 miR ⁃216b ⁃5p 来增强转化生长因子(transforming growth factor,TGF)⁃β1诱导的增殖反应[13]

  • 本课题前期研究已发现,用IL⁃17刺激NSCLC细胞(H1299和PC9)3 h,LINC01518的表达水平已明显提高,然而IL⁃17是否通过LINC01518引起NSCLC细胞增殖,目前尚不知晓,故本研究进行了探讨。

  • 1 材料和方法

  • 1.1 材料

  • 人正常支气管上皮细胞系 BEAS⁃2B 和 NSCLC 细胞系PC9、H1299和H1975(武汉中国典型培养物保藏中心)。胎牛血清(南京维森特公司),IL⁃17RA 抗体(武汉ABclonal公司),Lipofectamine2000(赛默飞公司,美国),人IL⁃17(R&D Systems公司,美国); CCK⁃8试剂盒(南京诺唯赞公司),EdU试剂盒(广州锐博公司);pcDNA3.1/LINC01518(合肥通用公司), shLINC01518(上海吉玛公司);ABI Simpli Amp PCR 热循环仪和 ABI Step One Plus 定量 PCR 仪(赛默飞公司,美国),倒置显微镜 IX73(奥林巴斯公司,日本)。

  • 1.2 方法

  • 1.2.1 细胞系的培养与传代

  • 将 PC9、H1299、H1975 和 BEAS ⁃ 2B 接种于含 10%胎牛血清的DMEM完全培养液中,于37℃、5% CO2培养 48 h。当细胞融合度达 90%时按 1∶5 比例进行传代。

  • 1.2.2 Western blot

  • 将细胞裂解物离心3 min,取20 μL蛋白行SDS⁃ PAGE 电泳。先 45 V 恒压浓缩再 120 V 电泳 2 h, 0.3A 恒流湿转 120 min。PVDF 膜用脱脂牛奶封闭 2 h后加相应一抗4℃孵育过夜,漂洗后用HRP标记的二抗孵育1 h。最后行ECL化学发光试剂检测。

  • 1.2.3 RT⁃PCR

  • 用IL⁃17分别刺激PC9和H1299细胞,TRIzol提取细胞的总 RNA,逆转录为 cDNA(留作模板)。同时查阅相关文献[7-18],并从前期芯片结果(即上调2倍以上的 LncRNA 459 个)中挑选出上调较高且已报告有促增殖效应的10个LncRNA进行RT⁃PCR检查验证。检测前 Premier 5 软件设计的引物由合肥通用生物科技有限公司合成,序列见表1。

  • RT ⁃ PCR 反应总体系为 25 μL,含 cDNA 模板 1 μL,Taq Master Mix 12.5 μL,上下游引物各 1 μL,灭菌双蒸水补足至总体积。用ABI Simpli Amp PCR 热循环仪进行PCR反应,程序为:95℃ 15 min变性, 55℃ 30 s退火,72℃ 30 s延伸;共35个循环。

  • 1.2.4 Real⁃time PCR

  • 同上设计合成 LINC01518、LINC00265 和 PSMA3⁃AS1引物(序列见表1)。Real⁃time PCR反应体系为20 μL,含cDNA模板1 μL,SYBR Green Master Mix 10 μL,上下游引物各 0.4 μL,双蒸水补足总体积。用 ABI Step One Plus 定量 PCR 仪进行扩增反应,程序为:预变性,95℃ 5 min;循环反应,95℃ 10 s, 60℃ 30 s,共40个循环;熔解曲线,95℃ 15 s、60℃ 60 s、95℃ 15 s。数据以β⁃actin作为内参,2-ΔΔCT法计算相对表达量,Graphpad Prism 5进行半定量分析。

  • 1.2.5 质粒的构建与鉴定

  • 过表达的 pcDNA3.1/LINC01518 和小干扰的 shLINC01518 质粒(含 shLINC01518⁃1~3)分别由合肥通用生物和上海吉玛制药技术有限公司构建。 shLINC01518⁃1:5′⁃CCCGTACCTAGTTCTTAAA⁃3′; shLINC01518 ⁃ 2:5′ ⁃ ATGCCAACTTATCTGGCTA ⁃ AT⁃3′;shLINC01518⁃3:5′⁃TGGCCTAGATCATTAA⁃ CATAT⁃3′。测序鉴定证实,上述质粒均构建成功。

  • 将 pcDNA3.1/LINC01518 用 Lipofectamine2000转染H1299细胞48 h,在明确转染效率可达70%以上后进行实验分组:①pcDNA3.1 组;②pcDNA3.1/ LINC01518组。此外,采用同前方法将shLINC01518 质粒转染细胞后48 h(效率可达80%)再用IL⁃17刺激 3 h。实验分组:①shCTR+IL⁃17组;②shLINC01518⁃1+ IL⁃17组;③shLINC01518⁃2+IL⁃17组;④shLINC01518⁃3 +IL⁃17 组。提取前述不同处理细胞的总 RNA,行 Real⁃time PCR检查。

  • 表1 PCR引物

  • Table1 Primers for PCR

  • 1.2.6 CCK⁃8实验

  • IL⁃17 刺激 H1299 和 PC9 细胞。实验分组:① DMEM组:两种细胞仅用无血清DMEM培养;②IL⁃17 刺激组:取50 ng/mL的 IL⁃17(课题组以往研究确定该浓度为最佳刺激剂量)[6],每孔加150 μL,在刺激后不同时间加CCK⁃8溶液处理。

  • 过表达 LINC01518。实验分组:①pcDNA3.1 组;② pcDNA3.1/ LINC01518 组,将 pcDNA3.1 或 pcDNA3.1/LINC01518 转染 H1299 细胞 48 h。处理后加CCK⁃8溶液。

  • 沉默LINC01518后再加IL⁃17刺激。实验分组: ①shCTR 组,仅转染 shCTR 48 h;②shCTR + IL ⁃17 组,将 shCTR 转梁 H1299 细胞 48 h 再行 IL⁃17 刺激 48 h;③shLINC01518+IL⁃17 组,将 shLINC01518 转染H1299细胞48 h再行IL⁃17刺激48 h。最后每孔加10 μL CCK⁃8溶液,37℃孵育40 min,用酶标仪测定450 nm 处的吸光度值,以吸光度值表示细胞的增殖水平。

  • 1.2.7 EdU实验

  • 过表达或沉默 LINC01518 的分组处理同 1.2.6。每孔细胞先加100 μL 50 μmol/L EdU溶液孵育2 h,再用4%多聚甲醛孵育30 min,接着加2 mg/mL甘氨酸5 min,用0.5% Triton X⁃100 PBS脱色10 min,加Appllo染色液避光孵育30 min后用Hoechst 染色液避光孵育30 min。在荧光镜下观察拍摄5个随机选择的视野。

  • 1.2.8 克隆形成实验

  • 过表达或沉默 LINC01518 分组同 1.2.6 所述。当细胞培养 8 d 出现肉眼可见的克隆时终止培养。先弃去培养液,再用 PBS 浸洗,接着加甲醇固定 20 min后弃去,再加结晶紫染液染20 min,洗去染色液,干燥后用肉眼直接计数细胞克隆的形成数量。

  • 1.3 统计学方法

  • 采用 SPSS 22.0 软件进行统计学分析,使用 GraphPad Prism 8软件进行绘图。所有实验均独立重复3次,所得定量数据以均数±标准差(x-±s)进行统计描述。Student’s t 检验用于比较两组间差异; 多组间差异比较采用单因素方差分析,两两比较采用 SNK 检验,明确对照组的比较采用 Dunnett⁃t 检验,P <0.05为差异有统计学意义。

  • 2 结果

  • 2.1 3种NSCLC细胞系均有IL⁃17RA的明显表达

  • Western blot 检测NSCLC细胞表面的IL⁃17RA,结果显示,这些细胞均有IL⁃17RA的表达,H1299和 PC9细胞表达较多(图1A、B)。

  • 2.2 IL⁃17刺激H1299和PC9细胞不同时间可引起细胞的增殖

  • 在证实了 H1299 和 PC9 细胞可表达 IL ⁃17RA 后,为了确定 IL⁃17 刺激能诱导细胞增殖,用 IL⁃17 (50 ng/mL)刺激细胞,在刺激0、24、48、72、96 h时行 CCK⁃8 实验检测细胞增殖水平,发现 IL⁃17 刺激后24 h,两种细胞的增殖水平均已升高,48 h以后均较显著(图1C、D)。

  • 2.3 IL ⁃17 刺激 H1299 和 PC9 细胞后能显著升高 LINC01518的表达水平

  • 为了验证IL⁃17刺激H1299和PC9细胞后上调的 LncRNA,从前期 LncRNA 芯片上调 2 倍以上的 459 个 LncRNA 中,选取了 10 个文献已报道有促增殖效应的LncRNA[7-18],先行RT⁃PCR验证。结果显示,IL ⁃ 17 刺激两细胞后不同时间,LINC01518、 LINC00265 和 PSMA3 ⁃ AS1 表达升高明显(图2A~D)。Real⁃time PCR进一步检查前述3个升高的 LncRNA 发现,IL⁃17 刺激 2 h 和 3 h 时,LINC01518、 LINC00265 及 PSMA3 ⁃ AS1 均显著上升,其中 LINC01518升高得更为明显,且以H1299细胞较为显著(图2E、F),故此后选择H1299细胞继续开展实验。

  • 图1 NSCLC细胞系IL⁃17RA表达和IL⁃17刺激H1299和PC9细胞不同时间后细胞增殖水平的变化

  • Figure1 The expression of IL⁃17RA in NSCLC cell lines and the changes of H1299 and PC9 cell proliferation stimulated with IL⁃I7 for different times

  • 2.4 LINC01518过表达和小干扰质粒的鉴定、表达及干扰效率的验证

  • 为了探讨 LINC01518 在 IL⁃17 诱导 H1299 细胞增殖中的作用,先构建了LINC01518过表达和小干扰质粒。测序鉴定成功后(图3A、B),将pcDNA3.1/ LINC01518 质粒转染 H1299 细胞 48 h,行 Real⁃time PCR检测,结果显示转染pcDNA3.1/LINC01518的细胞中,LINC01518 的表达显著增加(图3C)。此外,转染 shLINC01518⁃1~3 小干扰质粒 48 h 再行 IL⁃17 刺激 3 h 发现,shLINC01518⁃1 能显著抑制 IL⁃17 上调的LINC01518的表达(图3D),提示shLINC01518⁃1 沉默效率最佳,故选取 shLINC01518⁃1 进行后续实验,并统一命名为shLINC01518。

  • 2.5 LINC01518 基因表达能促进 IL ⁃ 17 诱导的 H1299细胞增殖

  • 为了进一步检查LINC01518对H1299细胞增殖的影响,在细胞转染 pcDNA3.1/LINC01518 质粒和 shLINC01518质粒后进行了相应的实验。CCK⁃8实验表明,pcDNA3.1/LINC01518 转染组,48 h 时细胞增殖水平明显升高,而沉默 LINC01518 基因后由 IL⁃17诱导的细胞增殖未见增加(图4A)。同时,EdU 实验显示,pcDNA3.1/LINC01518 转染 48 h 时,进入增殖期的细胞数明显增多,而沉默shLINC01518基因后,由 IL⁃17 诱导进入增殖期的细胞数未见明显升高(图4B、C)。此外,细胞克隆形成实验发现,过表达LINC01518基因8 d时,细胞克隆数明显增加,而沉默LINC01518基因后8 d,由IL⁃17诱导的细胞克隆形成数则未见增多(图4D、E)。上述结果表明, IL⁃17刺激H1299细胞后可通过上调LINC01518的表达促进H1299细胞的增殖。

  • 3 讨论

  • 本课题组以往研究表明,NSCLC癌组织中IL⁃17 表达明显升高,且用 IL⁃17 刺激 NSCLC 细胞后能诱导其增殖,相应机制涉及 HMGA1 上调可致 cyclin D1基因的转录与表达[6]。不过,由于NSCLC细胞增殖的机制较为复杂,故深入研究癌细胞异常增殖的原因及其分子调控机制十分必要。

  • 已知细胞因子刺激细胞发挥效应的前提是其细胞表面含有相应的受体[6],故本研究一开始就检测了 NSCLC 细胞系 IL⁃17RA 的表达水平。在证实了 PC9、H1299和H1975细胞系IL⁃17RA的表达均高于正常支气管上皮细胞BEAS⁃2B后,又选择了IL⁃17RA 表达较高的PC9和H1299细胞,用定量IL⁃17刺激不同时间行 CCK⁃8 实验,确证两种细胞受 IL⁃17 刺激 24 h 时,增殖水平已明显升高,48 h以后更加显著。

  • 为了探讨 IL⁃17 刺激 NSCLC 细胞诱导其增殖的可能机制,本课题组前期已用 LncRNA 芯片筛查了 IL⁃17 刺激 3 h 后 H1299 和 PC9 细胞差异表达的LncRNA。结果发现,上调≥2 倍的 LncRNA 共有 459 个。查阅相关文献[7-18],并结合芯片中上调较高 (≥3 倍)的 LncRNA,从中选取了10条与细胞增殖有关的LncRNA,先用RT⁃PCR测定IL⁃17刺激3 h时两种细胞mRNA的水平,在筛出了LINC01518、LINC00265 和 PSMA3⁃AS1 升高显著后,又用 Real⁃time PCR 进一步确定前述 3 个表达水平升高的 LncRNA, LINC01518的上调倍数既明显又相对稳定。

  • 图2 IL⁃17刺激H1299和PC9细胞后不同时间LncRNA芯片上调明显的10个LncRNA的表达验证

  • Figure2 The expression validation of 10 up⁃regulated lncRNAs from the microarray in IL⁃17⁃stimulated H1299 and PC9 cells at different times

  • 图3 LINC01518过表达质粒的表达验证和shLINC01518质粒沉默效率的检测

  • Figure3 The determination of pcDNA3.1/LINC01518 expression and screening of most effective shRNA for silencing LINC01518

  • 图4 过表达和沉默LINC01518基因对H1299细胞增殖的影响

  • Figure4 The effects of overexpressing and silencing LINC01518 gene on the proliferation of H1299 cells

  • LINC01518 位于人染色体 10q11.21,其细胞定位和功能研究发现,细胞核中的LncRNA 多与染色质修饰、转录调节等过程相关,而细胞质中的LncRNA 则可作为竞争性内源 RNA(competing endogenous RNA,ceRNA)结合相应的miRNA,进而在转录后能间接调控靶基因[8-1012-1315-18]。研究发现,LINC01518 在食管癌细胞中的过量表达能通过 ceRNA 结合 miR⁃1⁃3p促进细胞增殖[11-12]。然而LINC01518是否参与IL⁃17诱导的NSCLC细胞增殖,目前尚无文献报道。

  • 本研究构建了 LINC01518 过表达(pcDNA3.1/ LINC01518)和 shLINC01518 小干扰质粒。在验证上述质粒构建成功后,分别将这些质粒转染H1299 细胞(转染 shLINC01518 质粒的细胞再行 IL⁃17 刺激)。结果显示,过表达LINC01518能显著提高细胞的增殖水平,而敲低LINC01518基因后由IL⁃17诱导的细胞增殖未见明显增加。这一结果提示,LINC01518 的表达可正向提升 IL⁃17 诱导 H1299 细胞增殖的能力。

  • LncRNA调控细胞生物学行为的机制之一是其能吸附miRNA影响靶基因的表达,故其在肿瘤等疾病的发生与发展中发挥了重要作用[8-1012-1315-18]。由于 LINC01518 可竞争结合某些 miRNA 来影响细胞的某些病变[12-13],故设想在NSCLC发病和进展过程中,LINC01518的表达上调或许也能通过结合相应的miRNA促进IL⁃17诱导的肿瘤细胞增殖反应。不过,这一推测还有待后续进一步对LINC01518的细胞定位和功能研究等加以验证。

  • 参考文献

    • [1] HERBST R S,MORGENSZTERN D,BOSHOFF C.The biology and management of non small cell lung cancer [J].Nature,2018,553(7689):446-454

    • [2] RICE S J,LIU X,ZHANG J H,et al.Advanced NSCLC patients with high IL ⁃ 6 levels have altered peripheral T cell population and signaling[J].Lung Cancer Amster⁃ dam Neth,2019,131:58-61

    • [3] KWIECIEN I,STELMASZCZYK⁃EMMEL A,POLUBIEC⁃ KOWNACKA M,et al.Elevated regulatory T cells,sur⁃ face and intracellular CTLA⁃4 expression and interleukin⁃ 17 in the lung cancer microenvironment in humans[J].Cancer Immunol Immunother,2017,66(2):161-170

    • [4] WANG B,ZHAO C H,SUN G,et al.IL ⁃17 induces the proliferation and migration of glioma cells through the ac⁃ tivation of PI3K/Akt1/NF⁃κB⁃p65[J].Cancer Lett,2019,447:93-104

    • [5] XIANG T,LONG H,HE L,et al.Interleukin⁃17 produced by tumor microenvironment promotes self ⁃ renewal of CD133 + cancer stem⁃like cells in ovarian cancer[J].On⁃ cogene,2015,34(2):165-176

    • [6] ZHAO C,LI Y,ZHANG W,et al.IL17 induces NSCLC A549 cell proliferation via the upregulation of HMGA1,resulting in an increased cyclin D1 expression[J].Int J Oncol,2018,52(5):1579-1592

    • [7] GE B H,ZHANG X,ZHOU W,et al.LINC00265 promotes metastasis and progression of hepatocellular carcinoma by interacting with E2F1 at the promoter of CDK2[J].Cell J,2022,24(6):294-301

    • [8] ZHENG G L,LIU Y L,YAN Z X,et al.Elevated LOXL2 expression by LINC01347/miR⁃328⁃5p axis contributes to 5 ⁃ FU chemotherapy resistance of colorectal cancer[J].Am J Cancer Res,2021,11(4):1572-1585

    • [9] CHENG G,LI Y,LIU Z,et al.LncRNA PSMA3⁃AS1 pro⁃ motes the progression of non ⁃ small cell lung cancer through targeting miR ⁃ 17 ⁃ 5p/PD ⁃ L1[J].Adv Clin Exp Med,2021,30(10):1043-1050

    • [10] XING Z,ZHANG Z,GAO Y,et al.The lncRNA LINC01194/miR ⁃ 486 ⁃ 5p axis facilitates malignancy in non⁃small cell lung cancer via regulating CDK4[J].Onco Targets Ther,2020,13:3151-3163

    • [11] WANG W W,WEI C G,LI P,et al.Integrative analysis of mRNA and lncRNA profiles identified pathogenetic lncRNAs in esophageal squamous cell carcinoma[J].Gene,2018,661:169-175

    • [12] ZHANG D H,ZHANG H F,WANG X L,et al.LINC01518 knockdown inhibits tumorigenicity by suppression of PIK3CA/Akt pathway in oesophageal squamous cell carci⁃ noma[J].Artif Cells Nanomed Biotechnol,2019,47(1):4284-4292

    • [13] KONG N,BAO Y,ZHAO H,et al.Long noncoding RNA LINC01518 modulates proliferation and migration in TGF⁃ β1 ⁃treated human tenon capsule fibroblast cells through the regulation of hsa ⁃miR ⁃ 216b ⁃5p[J].Neuromolecular Med,2022,24(2):88-96

    • [14] YOJIRO K,NATSUMI M,TAKAHIRO W,et al.OIP5 ⁃ AS1 promotes proliferation of non⁃small⁃cell lung cancer and head and neck squamous cell carcinoma cells[J].Cancer Genom Proteom,2021,18(4):543-548

    • [15] PENG X,WEI F,HU X.Long noncoding RNA DLGAP1⁃ AS1 promotes cell proliferation in hepatocellular carcino⁃ ma via sequestering miR ⁃ 486 ⁃ 5p[J].J Cell Biochem,2020,121(2):1953-1962

    • [16] CHEN W H,WANG L F,LI X Y,et al.LncRNA SNHG17 regulates cell proliferation and invasion by targeting miR⁃ 338⁃3p/SOX4 axis in esophageal squamous cell carcinoma [J].Cell Death Dis,2021,12(9):806

    • [17] WANG J,YU Z Y,WANG J,et al.LncRNA NUTM2A ⁃ AS1 positively modulates TET1 and HIF ⁃ 1A to enhance gastric cancer tumorigenesis and drug resistance by spong⁃ ing miR⁃376a[J].Cancer Med,2020,9(24):9499-9510

    • [18] YANG Y,YU Q,LI B,et al.BBOX1⁃AS1 accelerates gas⁃ tric cancer proliferation by sponging miR⁃3940⁃3p to up⁃ regulate BIRC5 expression[J].Dig Dis Sci,2021,66(4):1054-1062

  • 参考文献

    • [1] HERBST R S,MORGENSZTERN D,BOSHOFF C.The biology and management of non small cell lung cancer [J].Nature,2018,553(7689):446-454

    • [2] RICE S J,LIU X,ZHANG J H,et al.Advanced NSCLC patients with high IL ⁃ 6 levels have altered peripheral T cell population and signaling[J].Lung Cancer Amster⁃ dam Neth,2019,131:58-61

    • [3] KWIECIEN I,STELMASZCZYK⁃EMMEL A,POLUBIEC⁃ KOWNACKA M,et al.Elevated regulatory T cells,sur⁃ face and intracellular CTLA⁃4 expression and interleukin⁃ 17 in the lung cancer microenvironment in humans[J].Cancer Immunol Immunother,2017,66(2):161-170

    • [4] WANG B,ZHAO C H,SUN G,et al.IL ⁃17 induces the proliferation and migration of glioma cells through the ac⁃ tivation of PI3K/Akt1/NF⁃κB⁃p65[J].Cancer Lett,2019,447:93-104

    • [5] XIANG T,LONG H,HE L,et al.Interleukin⁃17 produced by tumor microenvironment promotes self ⁃ renewal of CD133 + cancer stem⁃like cells in ovarian cancer[J].On⁃ cogene,2015,34(2):165-176

    • [6] ZHAO C,LI Y,ZHANG W,et al.IL17 induces NSCLC A549 cell proliferation via the upregulation of HMGA1,resulting in an increased cyclin D1 expression[J].Int J Oncol,2018,52(5):1579-1592

    • [7] GE B H,ZHANG X,ZHOU W,et al.LINC00265 promotes metastasis and progression of hepatocellular carcinoma by interacting with E2F1 at the promoter of CDK2[J].Cell J,2022,24(6):294-301

    • [8] ZHENG G L,LIU Y L,YAN Z X,et al.Elevated LOXL2 expression by LINC01347/miR⁃328⁃5p axis contributes to 5 ⁃ FU chemotherapy resistance of colorectal cancer[J].Am J Cancer Res,2021,11(4):1572-1585

    • [9] CHENG G,LI Y,LIU Z,et al.LncRNA PSMA3⁃AS1 pro⁃ motes the progression of non ⁃ small cell lung cancer through targeting miR ⁃ 17 ⁃ 5p/PD ⁃ L1[J].Adv Clin Exp Med,2021,30(10):1043-1050

    • [10] XING Z,ZHANG Z,GAO Y,et al.The lncRNA LINC01194/miR ⁃ 486 ⁃ 5p axis facilitates malignancy in non⁃small cell lung cancer via regulating CDK4[J].Onco Targets Ther,2020,13:3151-3163

    • [11] WANG W W,WEI C G,LI P,et al.Integrative analysis of mRNA and lncRNA profiles identified pathogenetic lncRNAs in esophageal squamous cell carcinoma[J].Gene,2018,661:169-175

    • [12] ZHANG D H,ZHANG H F,WANG X L,et al.LINC01518 knockdown inhibits tumorigenicity by suppression of PIK3CA/Akt pathway in oesophageal squamous cell carci⁃ noma[J].Artif Cells Nanomed Biotechnol,2019,47(1):4284-4292

    • [13] KONG N,BAO Y,ZHAO H,et al.Long noncoding RNA LINC01518 modulates proliferation and migration in TGF⁃ β1 ⁃treated human tenon capsule fibroblast cells through the regulation of hsa ⁃miR ⁃ 216b ⁃5p[J].Neuromolecular Med,2022,24(2):88-96

    • [14] YOJIRO K,NATSUMI M,TAKAHIRO W,et al.OIP5 ⁃ AS1 promotes proliferation of non⁃small⁃cell lung cancer and head and neck squamous cell carcinoma cells[J].Cancer Genom Proteom,2021,18(4):543-548

    • [15] PENG X,WEI F,HU X.Long noncoding RNA DLGAP1⁃ AS1 promotes cell proliferation in hepatocellular carcino⁃ ma via sequestering miR ⁃ 486 ⁃ 5p[J].J Cell Biochem,2020,121(2):1953-1962

    • [16] CHEN W H,WANG L F,LI X Y,et al.LncRNA SNHG17 regulates cell proliferation and invasion by targeting miR⁃ 338⁃3p/SOX4 axis in esophageal squamous cell carcinoma [J].Cell Death Dis,2021,12(9):806

    • [17] WANG J,YU Z Y,WANG J,et al.LncRNA NUTM2A ⁃ AS1 positively modulates TET1 and HIF ⁃ 1A to enhance gastric cancer tumorigenesis and drug resistance by spong⁃ ing miR⁃376a[J].Cancer Med,2020,9(24):9499-9510

    • [18] YANG Y,YU Q,LI B,et al.BBOX1⁃AS1 accelerates gas⁃ tric cancer proliferation by sponging miR⁃3940⁃3p to up⁃ regulate BIRC5 expression[J].Dig Dis Sci,2021,66(4):1054-1062

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