[1]才 昊,韩 亮,李晓飞.LncRNA XIST通过调控miR-186-5p促进非小细胞肺癌的增殖和侵袭实验研究*[J].陕西医学杂志,2020,49(7):781-787803.[doi:DOI:10.3969/j.issn.10007377.2020.07.004]
　CAI Hao,HAN Liang,LI Xiaofei..LncRNA XIST promotes proliferation and invasion of non-small cell lung cancer by regulating miR-186-5p[J].,2020,49(7):781-787803.[doi:DOI:10.3969/j.issn.10007377.2020.07.004]
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LncRNA XIST通过调控miR-186-5p促进非小细胞肺癌的增殖和侵袭实验研究*
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
49
期数:
2020年7期
页码:
781-787803
栏目:
基础研究
出版日期:
2020-07-05

文章信息/Info

Title:
LncRNA XIST promotes proliferation and invasion of non-small cell lung cancer by regulating miR-186-5p
作者:
才 昊; 韩 亮; 李晓飞
锦州医科大学附属第三医院胸外科(锦州 121000)
Author(s):
CAI Hao; HAN Liang; LI Xiaofei.
Department of Thoracic Surgery,the Third Affiliated Hospital of Jinzhou Medical University(Jinzhou 121000)
关键词:
LncRNA XIST;  miR-186-5p;  非小细胞肺癌;  增殖;  侵袭;  作用机制
Keywords:
LncRNA XIST;  MiR-186-5p;  Non-small cell lung cancer;  Proliferation;  Invasion;  Mechanism of action
分类号:
R734.2
DOI:
DOI:10.3969/j.issn.10007377.2020.07.004
文献标志码:
A
摘要:
目的:探究LncRNA XIST通过调控miR-186-5p促进非小细胞肺癌增殖和侵袭的作用机制。方法:选用非小细胞肺癌(NSCLC)细胞系A549、H1299、Calu-3及人支气管上皮细胞系BEAS-2B进行研究。采用RT-PCR检测NSCLC细胞系中LncRNA XIST表达情况。构建XIST siRNA转染敲降 XIST表达,采用MTT法、Transwell实验及Western blot法验证其对NSCLC细胞增殖、侵袭及相关蛋白表达的影响。构建包含miR-186-5p结合位点的pmir GLO-XIST-Wt和pmir GLO-XIST-Mut基因片段报告载体,采用荧光素基因实验进行两者结合验证,并利用RIP实验及营救实验检测验证XIST和miR-186-5p靶向调控关系。结果:NSCLC细胞系A549、H1299、Calu-3中LncRNA XIST表达水平均显著高于人支气管上皮细胞系BEAS-2B(P<0.05)。敲降XIST表达后A549、H1299细胞中LncRNA XIST表达显著降低,miR-186-5p表达显著增高(P<0.05)。过表达miR-186-5p可显著降低A549、H1299细胞中XIST表达。敲降XIST显著抑制A549、H1299细胞增殖、侵袭能力,降低CCND1、PCNA、MMP-9、Bcl-2蛋白表达水平(P<0.05)。XIST是miR-186-5p的靶向基因。XIST与miR-186-5p存在于相同RISC复合物中,miR-186-5p以Ago2依赖的方式调控XIST。过表达miR-186-5p后A549细胞增殖、侵袭能力显著降低,逆转XIST过表达后促进A549细胞增殖、侵袭。结论:LncRNA XIST在非小细胞肺癌中高表达,LncRNA XIST通过调控miR-186-5p进而抑制XIST表达影响NSCLC的生物学行为。
Abstract:
Objective:To explore the mechanism of LncRNA XIST promoting proliferation and invasion of non-small cell lung cancer(NSCLC)by regulating miR-186-5p. Methods:NSCLC cell lines A549,H1299,Calu-3 and human bronchial epithelial cell line BEAS-2B were selected for study. LncRNA XIST expression in NSCLC cell lines was detected by RT-PCR. XIST expression was knocked down by transfection of XIST siRNA,and its effect on proliferation,invasion and related protein expression of NSCLC cells was verified by MTT,Transwell experiment and Western blot. Wild type and mutant XIST gene fragment reporter vectors containing miR-186-5p binding sites were constructed,and the combination of the two was verified by the luciferin gene experiment,and the targeted regulatory relationship between XIST and miR-186-5p was verified by RIP experiment and rescue experiment. Results:LncRNA XIST expression levels in NSCLC cell lines A549,H1299 and Calu-3 were significantly higher than those in human bronchial epithelial cell line BEAS-2B(P<0.05). After knockdown of XIST expression,LncRNA XIST expression in A549 and H1299 cells was significantly decreased,and miR-186-5p expression was significantly increased(P<0.05). Overexpression of miR-186-5p significantly reduced XIST expression in A549 and H1299 cells. Knockdown XIST significantly inhibited the proliferation and invasion of A549 and H1299 cells,and reduced the expression levels of CCND1,PCNA,MMP-9 and Bcl-2 proteins(P<0.05). XIST is the target gene of miR-186-5p. XIST and miR-186-5p exist in the same RISC complex,and miR-186-5p regulates XIST in Ago2-dependent manner. Overexpression of miR-186-5p significantly inhibited the activity and invasion ability of A549 cells,and inhibited promoting role of XIST on proliferation and invasion of A549 cells. Conclusion:LncRNA XIST is highly expressed in NSCLC,and LncRNA XIST directly binds with miR-186-5p to affect proliferation and invasion of NSCLC by regulating miR-186-5p expression. XIST can be used as a new clinical therapeutic target for NSCLC.

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备注/Memo

备注/Memo:
*辽宁省自然科学基金资助项目(20180550458)
更新日期/Last Update: 2020-07-28