留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

circ_0091579/miR-330-3p/RNF126轴影响结直肠癌LoVo细胞的增殖、侵袭和凋亡

杨兴广 艾可为 杨继武 程伟 赵鹏举

杨兴广, 艾可为, 杨继武, 程伟, 赵鹏举. circ_0091579/miR-330-3p/RNF126轴影响结直肠癌LoVo细胞的增殖、侵袭和凋亡[J]. 中国肿瘤生物治疗杂志, 2021, 28(5): 460-468. doi: 10.3872/j.issn.1007-385x.2021.05.006
引用本文: 杨兴广, 艾可为, 杨继武, 程伟, 赵鹏举. circ_0091579/miR-330-3p/RNF126轴影响结直肠癌LoVo细胞的增殖、侵袭和凋亡[J]. 中国肿瘤生物治疗杂志, 2021, 28(5): 460-468. doi: 10.3872/j.issn.1007-385x.2021.05.006
YANG Xingguang, AI Kewei, YANG Jiwu, CHENG Wei, ZHAO Pengju. circ_0091579/miR-330-3p/RNF126 axis affects the proliferation, invasion and apoptosis of colorectal cancer LoVo cells[J]. Chin J Cancer Biother, 2021, 28(5): 460-468. doi: 10.3872/j.issn.1007-385x.2021.05.006
Citation: YANG Xingguang, AI Kewei, YANG Jiwu, CHENG Wei, ZHAO Pengju. circ_0091579/miR-330-3p/RNF126 axis affects the proliferation, invasion and apoptosis of colorectal cancer LoVo cells[J]. Chin J Cancer Biother, 2021, 28(5): 460-468. doi: 10.3872/j.issn.1007-385x.2021.05.006

circ_0091579/miR-330-3p/RNF126轴影响结直肠癌LoVo细胞的增殖、侵袭和凋亡

doi: 10.3872/j.issn.1007-385x.2021.05.006
基金项目: 云南省教育厅科学研究基金资助项目(No. 2019J0767)
详细信息
    作者简介:

    杨兴广(1986–),男,硕士,主治医师,主要从事普外科相关疾病的临床和基础研究,E-mail:yingtian008133@163.com

    通讯作者:

    艾可为(AI Kewei,corresponding author),本科,主任医师,主要从事普外科及肿瘤相关疾病的研究,E-mail:13577880176@139.com

    杨继武(YANG Jiwu,co-corresponding author),本科,教授、主任医师,主要从事普外科相关疾病的研究,E-mail:yangjiwu64@126.com

  • 中图分类号: R735.3;R730.54

circ_0091579/miR-330-3p/RNF126 axis affects the proliferation, invasion and apoptosis of colorectal cancer LoVo cells

Funds: Project supported by the Scientific Research Fund of the Education Department of Yunnan Province (No. 2019J0767)
  • 摘要:   目的:  探讨环状RNA circ_0091579作为分子海绵吸附miR-330-3p介导环指蛋白126(ring finger protein 126,RNF126)对结直肠癌(colorectal cancer,CRC)细胞增殖、凋亡、侵袭的影响。  方法:  选取2019年2月至2020年5月在大理大学第一附属医院行手术治疗的60例CRC患者的癌组织和癌旁组织。构建circ_0091579、miR-330-3p的过表达或敲减的CRC LoVo细胞,qPCR检测CRC组织和细胞中circ_0091579、miR-330-3p和RNF126的表达;MTT、Transwell、流式细胞术分别检测细胞的增殖、侵袭、凋亡情况;生物信息学方法预测circ_0091579和miR-330-3p、miR-330-3p和RNF126的靶向关系并用双荧光素酶报告实验和RNA免疫沉淀实验验证。  结果:  CRC组织和多种细胞(HCT116、SW620、CW-2、LoVo细胞)中,circ_0091579和RNF126均高表达、miR-330-3p均低表达(均P<0.05)。敲减circ_0091579可以抑制LoVo细胞的增殖、侵袭而促进其凋亡(均P<0.05),但该影响在加入miR-330-3p-inhibitor后被逆转;过表达miR-330-3p使LoVo细胞增殖和侵袭能力减弱但凋亡程度加强(均P<0.05),该影响在加入RNF126后被抵消。circ_0091579、miR-330-3p和RNF126之间存在靶向作用关系。  结论:  circ_0091579通过miR-330-3p/RNF126轴促进LoVo细胞增殖、侵袭而抑制其凋亡。
  • 图  1  circ_0091579、miR-330-3p在CRC组织和细胞中的相对表达量

    Figure  1.  Relative expressions of circ_0091579 and miR-330-3p in CRC tissues and cells

    *P<0.05 vs NCM460 cells A: The expression level of miR-330-3p in different cancer tissues predicted by the dbDEMC website; B, C: circ_0091579 expression was increased in CRC tissues and cells; D, E: miR-330-3p expression was decreased in CRC tissues and cells; F: There is a negative correlation between the expression levels of circ_0091579 and miR-330-3p

    图  2  qPCR检测circ_0091579

    Figure  2.  Characteristics of circ_0091579 detected by qPCR

    A: circ_0091579 can not be digested by RNase R; B: Results of nucleus-cytoplasm separation

    图  3  circ_0091579和miR-330-3p靶向关系验证结果

    Figure  3.  Verification of the targeting relationship between circ_0091579 and miR-330-3p

    A: There were binding sites between circ_0091579 and miR-330-3p predicted by online website; B: The targeting relationship between circ_0091579 and miR-330-3p was confirmed by Dual luciferase reporter assay; C: The targeting relationship between circ_0091579 and miR-330-3p was confirmed by RIP assay

    图  4  circ_0091579和miR-330-3p的表达对LoVo细胞增殖、侵袭和凋亡的影响

    Figure  4.  Effects of circ_0091579 and miR-330-3p expression on the proliferation, invasion and apoptosis of LoVo cells

    *P<0.05 vs si-NC group, P<0.05 vs si-circ_0091579+anti-miR-NC group A: Knockdown of circ_0091579 inhibited the proliferation of LoVo cells detected by MTT assay; B: Knockdown of circ_0091579 inhibited LoVo cell invasion detected by Transwell assay (×100); C: Knockdown of circ_0091579 promoted the apoptosis of LoVo cells detected by Flow cytometry

    图  5  RNF126和miR-330-3p的靶向关系验证结果

    Figure  5.  Verification of the targeting relationship between RNF126 and miR-330-3p

    *P<0.05 vs NCM460 cell line A: RNF126 expression was up-regulated in cancer tissues predicted by UALCAN website; B, C: RNF126 expression was increased in CRC tissues and cells; D: Correlation between RNF126 and miR-330-3p; E: There were binding sites between RNF126 and miR-330-3p; F, G: The targeting relationship between RNF126 and miR-330-3p was confirmed by Dual luciferase reporter assay and RIP assay

    图  6  miR-330-3p和RNF126的表达对LoVo细胞增殖、侵袭和凋亡的影响

    Figure  6.  Effects of miR-330-3p and RNF126 expression on the proliferation, invasion and apoptosis of LoVo cells

    *P<0.05 vs miR-NC group, P<0.05 vs miR-330-3p-mimic+vector group A: Results of MTT experiment in each group of cells; B: Transwell assay results in each group(×100); C: Results of Flow cytometry

    图  7  circ_0091579/miR-330-3p/RNF126轴参与CRC进展并影响CRC细胞生物学行为的示意图

    Figure  7.  Schematic diagram of circ_0091579/miR-330-3p/ RNF126 axis participate in the progression of CRC and affect the biological characteristics of CRC cells

    表  1  引物序列

    Table  1.   Primer sequences

    TargetSequence(5’-3’)
    circ_0091579 F: TGAGCCAGTGGT CAGTCAAA
    R: GTGGAGTCAGGCTTGG GTAG
    miR-330-3p F: GCAGAGATTCCGTTGTCGT
    R: GCGAGCACAGAATTAATACGAC
    RNF126 F: AAAAGAATTCGCCGAGGCGTCGCCGCATCC
    R: AAAACTCGAGTCACGAGTTGCTTGTGGCGTTCTCG
    GAPDH F: CTTTGGTATCGTGGAAGGACTC
    R: GTAGAGGCAGGGATGATGTTCT
    U6 F: CTTCGGCAGCACATATACTA
    R: AACTGGTGTCGTGGAGTC
    下载: 导出CSV

    表  2  circ_0091579表达与CRC患者临床病理参数相关性分析

    Table  2.   Correlation between circ_0091579 expression and clinicopathological parameters of patients with CRC

    ParameterNcirc_0091579 expression (n)P
    LowHigh
    Total case 60 26 34
    Age (t/a)
    <65 28 9 19 0.1232
    ≥65 32 17 15
    Gender
    Male 20 7 13 0.4162
    Female 40 19 21
    TNM
    0-Ⅱ 22 17 5 0.0001
    Ⅲ-Ⅳ 38 9 29
    Tumor size(d/cm)
    <5 24 18 9 0.0015
    ≥5 36 8 25
    Lymph node metastasis
    Yes 49 22 27 0.7419
    No 11 4 7
    Differentiation grade
    Medium/Low 42 14 28 0.0238
    High 18 12 6
    下载: 导出CSV
  • [1] WU G H, XUE M, ZHAO Y J, et al. Long noncoding RNA ZEB1-AS1 acts as a sponge of miR-141-3p to inhibit cell proliferation in colorectal cancer[J]. Int J Med Sci, 2020, 17(11): 1589-1597. DOI: 10.7150/ijms.46698.
    [2] DU F, CAO T, XIE H H, et al. KRAS mutation-responsive miR-139-5p inhibits colorectal cancer progression and is repressed by Wnt Signaling[J/OL]. Theranostics, 2020, 10(16): 7335-7350[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7330859/. DOI: 10.7150/thno.45971.
    [3] WANG J P, LIU S D, WANG H, et al. Robust performance of a novel stool DNA test of methylated SDC2 for colorectal cancer detection: a multicenter clinical study[J/OL]. Clin Epigenetics, 2020, 12(1): 162[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7602331/. DOI: 10.1186/s13148-020-00954-x.
    [4] LIU R, SHEN L, LIN C, et al. MiR-1587 regulates DNA damage repair and the radiosensitivity of CRC cells via targeting LIG4[J]. Dose Response, 2020, 18(2): 1559325820936906. DOI: 10.1177/1559325820936906.
    [5] CHEN X Q, JIANG J, ZHAO Y X, et al. Circular RNA circNTRK2 facilitates the progression of esophageal squamous cell carcinoma through up-regulating NRIP1 expression via miR-140-3p[J/OL]. J Exp Clin Cancer Res, 2020, 39(1): 133[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7353745/. DOI: 10.1186/s13046-020-01640-9.
    [6] XU Q, DENG B, LI M L, et al. circRNA-UBAP2 promotes the proliferation and inhibits apoptosis of ovarian cancer though miR-382-5p/PRPF8 axis[J/OL]. J Ovarian Res, 2020, 13(1): 81[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7372761/. DOI: 10.1186/s13048-020-00685-w.
    [7] XU W G, ZHOU B, WU J, et al. Circular RNA hsa-circ-0007766 modulates the progression of Gastric Carcinoma via miR-1233-3p/GDF15 axis[J/OL]. Int J Med Sci, 2020, 17(11): 1569-1583[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7359391/. DOI: 10.7150/ijms.46261.
    [8] ZHANG L, DONG X Q, YAN B, et al. CircAGFG1 drives metastasis and stemness in colorectal cancer by modulating YY1/CTNNB1[J/OL]. Cell Death Dis, 2020, 11(7): 542[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7367849/. DOI: 10.1038/s41419-020-2707-6.
    [9] DU H, HE Z, FENG F, et al. Hsa_circ_0038646 promotes cell proliferation and migration in colorectal cancer via miR-331-3p/GRIK3[J]. Oncol Lett, 2020, 20(1): 266-274. DOI: 10.3892/ol.2020.11547.
    [10] NIU W Y, CHEN L, ZHANG P, et al. Circ_0091579 promotes proliferative ability and metastasis of liver cancer cells by regulating microRNA-490-3p[J]. Eur Rev Med Pharmacol Sci, 2019, 23(23): 10264-10273. DOI: 10.26355/eurrev_201912_19664.
    [11] HUANG Y G, SUN H, MA X, et al. HLA-F-AS1/miR-330-3p/PFN1 axis promotes colorectal cancer progression[J]. Life Sci, 2020, 254: 117180. DOI: 10.1016/j.lfs.2019.117180.
    [12] WANG Z H, QU H J, GONG W J, et al. Up-regulation and tumor-promoting role of SPHK1 were attenuated by miR-330-3p in gastric cancer[J]. IUBMB Life, 2018, 70(11): 1164-1176. DOI: 10.1002/iub.1934.
    [13] MIGITA K, MATSUMOTO S, WAKATSUKI K, et al. RNF126 as a marker of prognosis and proliferation of gastric cancer[J]. Anticancer Res, 2020, 40(3): 1367-1374. DOI: 10.21873/anticanres.14078.
    [14] SHAHZAD N, MUNIR T, JAVED M, et al. SHISA3, an antagonist of the Wnt/β-catenin signaling, is epigenetically silenced and its ectopic expression suppresses growth in breast cancer[J/OL]. PLoS One, 2020, 15(7): e0236192[2020-12-12]. https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0236192. DOI: 10.1371/journal.pone.0236192.
    [15] PAN Q H, FAN Y H, WANG Y Z, et al. Long noncoding RNA NNT-AS1 functions as an oncogene in breast cancer via repressing ZFP36 expression[J]. J Biol Regul Homeost Agents, 2020, 34(3): 795-805. DOI: 10.23812/20-100-a-13.
    [16] SHI J, LIU C, CHEN C, et al. Circular RNA circMBOAT2 promotes prostate cancer progression via a miR-1271-5p/mTOR axis[J]. Aging (Albany NY), 2020, 12(13): 13255-13280. DOI: 10.18632/aging.103432.
    [17] YIN H F, WANG Q, HUANG X T, et al. Circ_0032627 accelerates proliferation of gastric cancer cells through regulating miR-502-5p expression[J]. Eur Rev Med Pharmacol Sci, 2020, 24(12): 6674-6681. DOI: 10.26355/eurrev_202006_21654.
    [18] LIU Y H, LI H, YE X Y, et al. Hsa_circ_0000231 knockdown inhibits the glycolysis and progression of colorectal cancer cells by regulating miR-502-5p/MYO6 axis[J/OL]. World J Surg Oncol, 2020, 18(1): 255[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7526375/. DOI: 10.1186/s12957-020-02033-0.
    [19] WANG X M, TAO G, HUANG D H, et al. Circular RNA NOX4 promotes the development of colorectal cancer via the microRNA-485-5p/CKS1B axis[J]. Oncol Rep, 2020, 44(5): 2009-2020. DOI: 10.3892/or.2020.7758.
    [20] LI S, WENG J, SONG F, et al. Circular RNA circZNF566 promotes hepatocellular carcinoma progression by sponging miR-4738-3p and regulating TDO2 expression[J]. Cell Death Dis, 2020, 11(6): 452. DOI: 10.1038/s41419-020-2616-8.
    [21] WEI C H, ZHANG R G, CAI Q, et al. MicroRNA-330-3p promotes brain metastasis and epithelial-mesenchymal transition via GRIA3 in non-small cell lung cancer[J/OL]. Aging (Albany NY), 2019, 11(17): 6734-6761[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6756898/. DOI: 10.18632/aging.102201.
    [22] XIONG X, SHI Q, YANG X, et al. LINC00052 functions as a tumor suppressor through negatively modulating miR-330-3p in pancreatic cancer[J]. J Cell Physiol, 2019, 234(9): 15619-15626. DOI: 10.1002/jcp.28209.
    [23] JIN Z, JIA B, TAN L, et al. miR-330-3p suppresses liver cancer cell migration by targeting MAP2K1[J]. Oncol Lett, 2019, 18(1): 314-320. DOI: 10.3892/ol.2019.10280.
    [24] MANSOORI B, MOHAMMADI A, NAGHIZADEH S, et al. miR-330 suppresses EMT and induces apoptosis by downregulating HMGA2 in human colorectal cancer[J]. J Cell Physiol, 2020, 235(2): 920-931. DOI: 10.1002/jcp.29007.
    [25] YOKOO H, OHOKA N, NAITO M, et al. Design and synthesis of peptide-based chimeric molecules to induce degradation of the estrogen and androgen receptors[J]. Bioorg Med Chem, 2020, 28(15): 115595. DOI: 10.1016/j.bmc.2020.115595.
    [26] ALIX E, GODLEE C, CERNY O, et al. The tumour suppressor TMEM127 is a Nedd4-family E3 ligase adaptor required by Salmonella SteD to ubiquitinate and degrade MHC class Ⅱ molecules[J/OL]. Cell Host Microbe, 2020, 28(1): 54-68. e7[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7342019/. DOI: 10.1016/j.chom.2020.04.024.
    [27] KHAN S, HE Y, ZHANG X, et al. PROteolysis TArgeting Chimeras (PROTACs) as emerging anticancer therapeutics[J]. Oncogene, 2020, 39(26): 4909-4924. DOI: 10.1038/s41388-020-1336-y.
    [28] WANG L N, WANG X, ZHAO Y H, et al. E3 Ubiquitin ligase RNF126 regulates the progression of tongue cancer[J/OL]. Cancer Med, 2016, 5(8): 2043-2047[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4971932/. DOI: 10.1002/cam4.771.
    [29] YANG X S, PAN Y, QIU Z J, et al. RNF126 as a biomarker of a poor prognosis in invasive breast cancer and CHEK1 inhibitor efficacy in breast cancer cells[J/OL]. Clin Cancer Res, 2018, 24(7): 1629-1643[2020-12-12]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5884735/. DOI: 10.1158/1078-0432.CCR-17-2242.
  • 加载中
图(7) / 表(2)
计量
  • 文章访问数:  45
  • HTML全文浏览量:  25
  • PDF下载量:  1
  • 被引次数: 0
出版历程
  • 收稿日期:  2020-11-11
  • 修回日期:  2021-03-26
  • 网络出版日期:  2021-07-13
  • 刊出日期:  2021-05-25

目录

    /

    返回文章
    返回