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circ_0072088 通过调控 miR-545-3p/STAT3 轴促进非小细胞肺癌细胞的恶性生物学行为

项保利 王布 林卫佳 于亚楠

项保利, 王布, 林卫佳, 于亚楠. circ_0072088 通过调控 miR-545-3p/STAT3 轴促进非小细胞肺癌细胞的恶性生物学行为[J]. 中国肿瘤生物治疗杂志, 2021, 28(5): 469-476. doi: 10.3872/j.issn.1007-385x.2021.05.007
引用本文: 项保利, 王布, 林卫佳, 于亚楠. circ_0072088 通过调控 miR-545-3p/STAT3 轴促进非小细胞肺癌细胞的恶性生物学行为[J]. 中国肿瘤生物治疗杂志, 2021, 28(5): 469-476. doi: 10.3872/j.issn.1007-385x.2021.05.007
XIANG Baoli, WANG Bu, LIN Weijia, YU Yanan. circ_0072088 promotes the malignant biological behaviors of non-small cell lung cancer cells by regulating miR-545-3p/STAT3 axis[J]. Chin J Cancer Biother, 2021, 28(5): 469-476. doi: 10.3872/j.issn.1007-385x.2021.05.007
Citation: XIANG Baoli, WANG Bu, LIN Weijia, YU Yanan. circ_0072088 promotes the malignant biological behaviors of non-small cell lung cancer cells by regulating miR-545-3p/STAT3 axis[J]. Chin J Cancer Biother, 2021, 28(5): 469-476. doi: 10.3872/j.issn.1007-385x.2021.05.007

circ_0072088 通过调控 miR-545-3p/STAT3 轴促进非小细胞肺癌细胞的恶性生物学行为

doi: 10.3872/j.issn.1007-385x.2021.05.007
基金项目: 张家口市科学技术研究与发展计划资助项目(No.1911021D-3)
详细信息
    作者简介:

    项保利(1979–),男,硕士,副主任医师,主要从事呼吸介入治疗、肺癌诊治和肿瘤分子生物学研究工作,E-mail:liao3254@163.com

    通讯作者:

    王布(WANG Bu,corresponding author),男,硕士,副主任医师,主要从事呼吸病学相关的研究和诊治工作,E-mail:289974726@qq.com

  • 中图分类号: R734.2;R730.2

circ_0072088 promotes the malignant biological behaviors of non-small cell lung cancer cells by regulating miR-545-3p/STAT3 axis

Funds: Project supported by the Science and Technology Research and Development Plan Project of Zhangjiakou City (No.1911021D-3)
  • 摘要:   目的:  探究环状 RNA(circular RNA, circRNA)0072088 在非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞中的生物学功能及其作用机制。  方法:  在公共基因芯片数据库 Gene Expression Omnibus(GEO)中下载 GSE101684 数据集,通过 GEO2R 分析得到差异基因。通过qPCR检测NSCLC 细胞 H165、H358、H460、H226 和 A549 细胞中 circ_0072088 的表达水平,随后采用CCK-8法和 Transwell 小室法分别检测 circ_0072088 对 NSCLC 细胞增殖、迁移和侵袭的作用。通过 CircInteractome 和 TargetScan 数据库预测 circ_0072088 与miR-545-3p、miR-545-3p 与STAT3之间的靶向关系,并通过双荧光素酶报告基因实验和 RNA 结合蛋白免疫沉淀(RNA binding protein immunoprecipitation,RIP)实验验证 circ_0072088、miR-545-3p 与 STAT3 之间的靶向关系。  结果:  circ_0072088 在 NSCLC 细胞系中的表达均显著上调(均P<0.05)。过表达 circ_0072088 促进了 NSCLC 细胞的增殖、侵袭和迁移(均P<0.05);敲低 circ_0072088 抑制了 NSCLC 细胞的增殖、侵袭和迁移(均 P<0.05)。miR-545-3p 是 circ_0072088 的下游靶点,可以被 circ_0072088 吸附;STAT3 是 miR-545-3p 的靶基因,可以被 circ_0072088 间接正向调控。  结论:  Circ_0072088 通过调节 miR-545-3p /STAT3 轴促进 NSCLC 细胞的增殖和转移。
  • 图  1  circ_0072088 在 NSCLC 组织中的表达

    Figure  1.  The expression of circ_0072088 in NSCLC tissues

    A: Volcano map was used to show the differentially expressed circRNAs in NSCLC tissues (log2|Fold/change|>1, P<0.05); B: Heat map was used to show the differential expression of circ_0072088 in NSCLC tissues

    图  2  circ_0072088 在 NSCLC 细胞中的表达

    Figure  2.  Expression of circ_0072088 in NSCLC cells

    *P<0.05, **P<0.01vs BEAS-2B, Vector or si-NC group A: circ_0072088 expression in NSCLC cells was detected by qPCR; B: The subcellular localization of circ_0072088 was verified by qPCR; C: Transfection efficiency was detected by qPCR

    图  3  Circ_0072088 对 A549 和 H358 细胞增殖、迁移和侵袭的影响

    Figure  3.  The effects of circ_0072088 on cell viability, migration and invasion of A549 and H358 cells

    *P<0.05, **P<0.01 vs Vector or si-NC group A: Cell viability was detected by CCK-8 assay; B: Cell migration and invasion were detected by Transwell assay (×40)

    图  4  circ_0072088 靶向调控 miR-545-3p的表达

    Figure  4.  circ_0072088 regulated the expression of miR-545-3p

    *P<0.05, **P<0.01 A: The binding site between circ_0072088 and miR-545-3p; B, C: Dual luciferase reporter gene experiment and RIP experiment verified the targeting relationship between miR-545-3p and circ_0072088; D: The effect of over-expression or knockdown of circ_0072088 on the expression of miR-545-3p

    图  5  过表达circ_0072088和miR-545-3p 对A549细胞增殖、迁移和侵袭能力的影响

    Figure  5.  Effects of overexpression of circ_0072088 and miR-545-3p on the proliferation, migration and invasion of A549 cells

    *P<0.05, **P<0.01 A: Effects of transfection of circ_0072088 overexpression plasmid and miR-545-3p mimic on miR545-3p expression in A549 cells detected by qPCR; B: CCK-8 experiment detected the effect of overexpression of circ_0072088 and miR-545-3p on the proliferation of A549 cells; C, D: Transwell experiment detected the effect of overexpression of circ_0072088 and miR-545-3p on the migration and invasion of A549 cells

    图  6  circ_0072088 通过吸附 miR-545-3p 促进A549细胞中 STAT3 蛋白的表达

    Figure  6.  circ_0072088 promoted the protein expression of STAT3 in A549 cells by sponging miR-545-3p

    *P<0.05, **P<0.01 A: Binding site between MiR-545-3p and STAT3 predicted by the Targetscan database; B: The targeting relationship between miR-545-3p and STAT3 gene detected by Dual luciferase reporter gene experiment; C: The influence of circ_0072088 and miR-545-3p on STAT3 expression

    表  1  引物序列

    Table  1.   Primer sequences

    TargetPrimer sequence
    circ_0072088 F: GACCTCTGTCAGCGAGTTCC
    R: CTGCCTGTAACTCCTCTTCAGT
    miR-545-3p F: TGGCTCAGTTCAGCAGGAAC
    R: Universal reverse primer
    GAPDH F: AGAAGGCTGGGGCTCATTTG
    R: AGGGGCCATCCACAGTCTTC
    U6 F: CTCGCTTCGGCAGCACA
    R: AACGCTTCACGAATT TGCGT
    下载: 导出CSV
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出版历程
  • 收稿日期:  2020-12-28
  • 修回日期:  2021-04-16
  • 网络出版日期:  2021-07-13
  • 刊出日期:  2021-05-25

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