«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《现代肿瘤医学》[ISSN:1672-4992/CN:61-1415/R]

期数:

2020年11期

页码:

1844-1850

栏目:

论著（基础研究）

出版日期:

2020-04-30

文章信息/Info

Title:

miR-224-5p targeted regulation KIF23 and affected the proliferation，migration and invasion of cervical cancer cells through the NF-κB signaling pathway

作者:

夏 莹¹; 周晓莉¹; 刘 杰¹; 吴绪峰²; 汪黎明²; 贺晓琪³

1.中国人民解放军中部战区总医院妇产科，湖北 武汉 430070;2.湖北省妇幼保健院妇产科，湖北 武汉 430070;3.华中科技大学同济医学院附属协和医院妇产科，湖北 武汉 430022

Author(s):

Xia Ying¹; Zhou Xiaoli¹; Liu Jie¹; Wu Xufeng²; Wang Liming²; He Xiaoqi³

1.Department of Obstetrics and Gynecology，General Hospital of the Central War Zone of the Chinese People's Liberation Army，Hubei Wuhan 430070，China；2.Obstetrics and Gynecology Department,Hubei Maternal and Child Health Hospital,Hubei Wuhan 430070，China；3.Department of Obstetrics and Gynecology，Union Hospital Affiliated to Tongji Medical College，Huazhong University of Science and Technology，Hubei Wuhan 430022，China.

关键词:

miR-224-5p; KIF23; NF-κB信号通路; 宫颈癌; 增殖

Keywords:

miR-224-5p; KIF23; NF-κB signaling pathway; cervical cancer; proliferation

分类号:

R737.33

DOI:

10.3969/j.issn.1672-4992.2020.11.008

文献标识码:

A

摘要:

目的：探索微小RNA-224-5p（miR-224-5p）对驱动蛋白超家族23（KIF23）的靶向关系及对宫颈癌细胞增殖、迁移和侵袭的影响。方法：采用实时定量PCR（qPCR）检测正常宫颈细胞（H8）和宫颈癌细胞（SiHa、HeLa、MS751、HT-3）中miR-224-5p和KIF23 mRNA表达，选择miR-224-5p表达量最低的HeLa细胞开展后续研究。在HeLa细胞中转染si-KIF23或miR-224-5p，探讨敲减KIF23或过表达miR-224-5p对HeLa细胞增殖、迁移和侵袭的影响。噻唑蓝（MTT）比色法检测细胞增殖，Transwell小室法检测细胞迁移和侵袭，蛋白质印迹法（Western Blot）检测KIF23、细胞周期蛋白D1（CyclinD1）、基质金属蛋白酶2（MMP2）、基质金属蛋白酶9（MMP9）和NF-κBp65蛋白水平；生物学信息预测和双荧光素酶报告基因实验分析miR-224-5p和KIF23之间的靶向关系；共转染si-KIF23和anti-miR-224-5p，观察miR-224-5p低表达对敲减KIF23诱导的HeLa细胞增殖、迁移、侵袭和NF-κB信号通路活化的影响。结果：miR-224-5p在SiHa、HeLa、MS751、HT-3细胞中表达下调（P<0.05），KIF23 mRNA和蛋白表达上调（P<0.05）。敲减KIF23或过表达miR-224-5p显著降低HeLa细胞存活率、细胞迁移数和侵袭数，并显著抑制CyclinD1、MMP2、MMP9蛋白水平（P<0.05）。另外，敲减KIF23显著降低细胞中NF-κBp65表达量（P<0.05）。miR-224-5p靶向调控KIF23表达。miR-224-5p低表达可以逆转敲减KIF23抑制HeLa增殖、迁移、侵袭及CyclinD1、MMP2、MMP9蛋白表达的作用，以及逆转敲减KIF23抑制NF-κB信号通路活化的作用。结论：miR-224-5p靶向调控KIF23并通过NF-κB信号通路抑制宫颈癌细胞增殖、迁移和侵袭。

Abstract:

Objective：To investigate the targeting relationship of microRNA-224-5p(miR-224-5p)on Kinesin family member 23(KIF23)and the effects on proliferation，migration and invasion of cervical cancer cells.Methods：Real-time quantitative PCR(qPCR)was used to detect the expression of miR-224-5p and KIF23 in normal cervical cells(H8)and cervical cancer cells(SiHa，HeLa，MS751，HT-3).The HeLa cells with the lowest expression of miR-224-5p were selected for further research.The effects of knocking down KIF23 or over-expressing miR-224-5p on the proliferation，migration and invasion of HeLa cells were studied by transfecting si-KIF23 or miR-224-5p into HeLa cells.MTT colorimetric assay was used to determined cell proliferation.Transwell cell assay was applied to detect cell migration and invasion.Western Blot was employed to inspect the levels of KIF23，CyclinD1，Matrix metalloproteinase 2(MMP2)，Matrix metalloproteinase 9(MMP9)and NF-κBp65.Bioinformatics prediction and dual luciferase reporter gene assay were used to confirm the targeting relationship between miR-224-5p and KIF23.si-KIF23 and anti-miR-224-5p were co-transfected to observe the effects of low expression of miR-224-5p on cell proliferation，migration，invasion and activation of NF-κB signaling pathway induced by knocking down KIF23.Results：The expression of miR-224-5p was down-regulated in SiHa，HeLa，MS751 and HT-3 cells(P<0.05)，and the expression of KIF23 was up-regulated(P<0.05).Knockdown of KIF23 or over-expression of miR-224-5p obviously reduced the survival rate，cell migration and invasion of HeLa cells，and significantly inhibited the levels of CyclinD1，MMP2 and MMP9 proteins(P<0.05).In addition，knockdown of KIF23 significantly decreased the expression of NF-κBp65(P<0.05).miR-224-5p targeted regulation the expression of KIF23.The low expression of miR-224-5p reversed the inhibition of KIF23 on HeLa proliferation，migration，invasion and the expression of CyclinD1，MMP2 and MMP9 proteins，as well as the inhibition of KIF23 on the activation of NF-κB signaling pathway.Conclusion：miR-224-5p targets KIF23 and inhibits the proliferation，migration and invasion of cervical cancer cells through NF-κB signaling pathway.

参考文献/References

［1］Bray F，Ferlay J，Soerjomataram I，et al.Global cancer statistics 2018：GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］.CA：A Cancer Journal for Clinicians，2018，68(6)：394-424.
［2］Cancer Genome Atlas Research Network.Integrated genomic and molecular characterization of cervical cancer［J］.Nature，2017，543(7645)：378-384.
［3］Small Jr W，Bacon MA，Bajaj A，et al.Cervical cancer：A global health crisis［J］.Cancer，2017，123(13)：2404-2412.
［4］Marth C，Landoni F，Mahner S，et al.Cervical cancer：ESMO Clinical practice guidelines for diagnosis，treatment and follow-up［J］.Annals of Oncology，2017，28(suppl_4)：iv72-iv83.
［5］Sathyanarayanan A，Chandrasekaran KS，Karunagaran D.microRNA-145 modulates epithelial-mesenchymal transition and suppresses proliferation，migration and invasion by targeting SIP1 in human cervical cancer cells［J］.Cellular Oncology，2017，40(2)：119-131.
［6］Wei WF，Zhou CF，Wu XG，et al.MicroRNA-221-3p，a TWIST2 target，promotes cervical cancer metastasis by directly targeting THBS2［J］.Cell Death & Disease，2017，8(12)：3220-3232.
［7］Granados-López A，Ruiz-Carrillo J，Servín-González L，et al.Use of mature miRNA strand selection in miRNAs families in cervical cancer development［J］.International Journal of Molecular Sciences，2017，18(2)：407-430.
［8］Zheng XL，Tang HW，Zhao XF，et al.Long non-coding RNA FTH1P3 facilitates uveal melanoma cell growth and invasion through miR-224-5p［J］.PloS One，2017，12(11)：e0184746.
［9］Li JC，Liu XM，Li CP，et al.miR-224-5p inhibits proliferation，migration，and invasion by targeting PIK3R3/AKT3 in uveal melanoma［J］.Journal of Cellular Biochemistry，2019，120(8)：12412-12421.
［10］Li W，Liang J，Zhang Z，et al.MicroRNA-329-3p targets MAPK1 to suppress cell proliferation，migration and invasion in cervical cancer［J］.Oncology Reports，2017，37(5)：2743-2750.
［11］Poudyal D，Herman A，Adelsberger JW，et al.A novel microRNA，hsa-miR-6852 differentially regulated by Interleukin-27 induces necrosis in cervical cancer cells by downregulating the FoxM1 expression［J］.Scientific Reports，2018，8(1)：900-912.
［12］Sun P，Shen Y，Gong JM，et al.A new microRNA expression signature for cervical cancer［J］.International Journal of Gynecologic Cancer，2017，27(2)：339-343.
［13］Gao C，Zhou C，Zhuang J，et al.MicroRNA expression in cervical cancer：novel diagnostic and prognostic biomarkers［J］.Journal of Cellular Biochemistry，2018，119(8)：7080-7090.
［14］Yang YL，Liu Y，Li GL，et al.microRNA-214 suppresses the growth of cervical cancer cells by targeting EZH2［J］.Oncology Letters，2018，16(5)：5679-5686.
［15］Hua FF，Liu SS，Zhu LH，et al.MiRNA-338-3p regulates cervical cancer cells proliferation by targeting MACC1 through MAPK signaling pathway［J］.Eur Rev Med Pharmacol Sci，2017，21(23)：5342-5352.
［16］Zhang Z，Wang J，Wang X，et al.MicroRNA-21 promotes proliferation，migration，and invasion of cervical cancer through targeting TIMP3［J］.Archives of Gynecology and Obstetrics，2018，297(2)：433-442.
［17］YE L，LI HJ，ZHANG F，et al.Meta-analysis of the expression and significance of KIF23 in non-small cell lung cancer based on Oncomine database［J］.Chinese Journal of Lung Cancer，2017，20(12)：822-826.［叶亮，李慧娟，张方，等.基于Oncomine数据库荟萃分析KIF23在非小细胞肺癌中的表达和意义［J］.中国肺癌杂志，2017，20(12)：822-826.］
［18］SONG SS，ZHANG Q，NIE YH，et al.Analysis of the expression and prognostic significance of KIF23 in breast cancer based on database［J］.Tianjin Medicine，2019，47(2)：127-130,228.［宋姗姗，张强，聂艳红，等.基于数据库分析KIF23在乳腺癌中的表达及预后意义［J］.天津医药，2019，47(2)：127-130,228.］
［19］LIU SJ，LIN Q，BAI MJ，et al.Effects of KIF23 gene silencing on proliferation，migration and invasion of hepatocellular carcinoma HepG2 cells［J］.Journal of Sun Yat-Sen University(Medical Sciences)，2018，39(1)：34-40.［刘素娟，林曲，柏明军，等.KIF23基因沉默对肝癌HepG2细胞增殖、迁移及侵袭的影响［J］.中山大学学报(医学版)，2018，39(1)：34-40.］
［20］Iltzsche F，Simon K，Stopp S，et al.An important role for Myb-MuvB and its target gene KIF23 in a mouse model of lung adenocarcinoma［J］.Oncogene，2017，36(1)：110-121.
［21］Vikberg AL，Vooder T，Lokk K，et al.Mutation analysis and copy number alterations of KIF23 in non-small-cell lung cancer exhibiting KIF23 over-expression［J］.Onco Targets and Therapy，2017(10）：4969-4979.
［22］Zhao C，Wang XB，Zhang YH，et al.MicroRNA-424 inhibits cell migration，invasion and epithelial-mesenchymal transition in human glioma by targeting KIF23 and functions as a novel prognostic predictor［J］.European Review for Medical and Pharmacological Sciences，2018，22(19)：6369-6378.
［23］Wolter P，Hanselmann S，Pattschull G，et al.Central spindle proteins and mitotic kinesins are direct transcriptional targets of MuvB，B-MYB and FOXM1 in breast cancer cell lines and are potential targets for therapy［J］.Oncotarget，2017，8(7)：11160-11172.
［24］Li XL，Ji YM，Song R，et al.KIF23 promotes gastric cancer by stimulating cell proliferation［J］.Disease Markers，2019（2019）：9751923.
［25］Lin CC，Kuo CL，Huang YP，et al.Demethoxycurcumin suppresses migration and invasion of human cervical cancer HeLa cells via inhibition of NF-κB pathways［J］.Anticancer Research，2018，38(5)：2761-2769.
［26］Chen HH，Chen SP，Zheng QL，et al.Genistein promotes proliferation of human cervical cancer cells through estrogen receptor-mediated PI3K/Akt-NF-κB pathway［J］.Journal of Cancer，2018，9(2)：288-295.

备注/Memo

备注/Memo:

湖北省自然科学基金项目（编号：WJ2015MA017）

常用功能

更新日期/Last Update: 1900-01-01