LncRNA PKMYT1AR promotes cancer stem cell maintenance in non-small cell lung cancer via activating Wnt signaling pathway.
LncRNA PKMYT1AR promotes cancer stem cell maintenance in non-small cell lung cancer via activating Wnt signaling pathway.
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LncRNA PKMYT1AR通过激活Wnt信号通路促进非小细胞肺癌干细胞维持
DOI：
10.1186/s12943-021-01469-6
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发表时间：
2021-12-02
期刊：
影响因子：
37.3
通讯作者：
Chen Y
中科院分区：
文献类型：
--
作者：
He Y;Jiang X;Duan L;Xiong Q;Yuan Y;Liu P;Jiang L;Shen Q;Zhao S;Yang C;Chen Y
背景非小细胞肺癌（Non-small cell lung cancer，NSCLC）是人类最常见的肺癌类型，具有多种病理特征。虽然许多信号通路和治疗靶点已被定义为在NSCLC中发挥重要作用，限制efficaciesachieved.MethodsBioinformatics方法被用来确定差异长非编码RNA在NSCLC中的表达。采用实时荧光定量RT-PCR检测PKMYT 1AR、miR-485- 5 p的表达。通过体外和体内功能分析研究PKMYT 1AR/miR-485- 5 p/PKMYT 1轴在调节细胞增殖、迁移和肿瘤生长中的功能作用。采用双荧光素酶报告基因分析、荧光原位杂交（FISH）、免疫印迹、免疫共沉淀等实验验证了其分子机制。（lncRNA，ENST 00000595422），称为PKMYT 1AR（PKMYT 1相关lncRNA），在非小细胞肺癌中由阴阳1（YY 1）因子诱导，尤其是在癌细胞系中（H358、H1975、H1299、H1650、A549和SPC-A1）与正常人支气管上皮细胞系（BEAS-2B）相比。我们发现PKMYT 1AR高表达与更差的临床结果相关，并且PKMYT 1AR的敲低抑制肿瘤细胞增殖、迁移和异种移植肿瘤形成能力。生物信息学分析和荧光素酶试验表明，PKMYT 1AR直接与miR-485- 5 p相互作用，减弱其下游致癌因子PKMYT 1（蛋白激酶，膜相关酪氨酸/苏氨酸1）在NSCLC中的抑制作用。此外，我们发现，与相互对照组相比，miR-485- 5 p在癌细胞系和从NSCLC患者分离的外周血血清中均下调。一致地，miR-485- 5 p的强制表达抑制肿瘤细胞的增殖和迁移能力。此外，我们提供的证据表明，PKMYT 1AR靶向反义寡核苷酸（阿索）显着抑制肿瘤生长在体内。结论PKMYT 1AR/ miR-485- 5 p/PKMYT 1轴可能通过抑制β-TrCP 1介导的β-catenin蛋白的泛素降解而促进NSCLC中肿瘤干细胞（cancer stem cells，CSC）的维持，从而促进肿瘤的发生。
BackgroundNon-small cell lung cancer (NSCLC) is the most common type of human lung cancers, which has diverse pathological features. Although many signaling pathways and therapeutic targets have been defined to play important roles in NSCLC, limiting efficacies have been achieved.MethodsBioinformatics methods were used to identify differential long non-coding RNA expression in NSCLC. Real-time RT-PCR experiments were used to examine the expression pattern of lncRNA PKMYT1AR, miR-485-5p. Both in vitro and in vivo functional assays were performed to investigate the functional role of PKMYT1AR/miR-485-5p/PKMYT1 axis on regulating cell proliferation, migration and tumor growth. Dual luciferase reporter assay, fluorescent in situ hybridization (FISH), immunoblot, co-immunoprecipitation experiments were used to verify the molecular mechanism.ResultHere, we identify a human-specific long non-coding RNA (lncRNA, ENST00000595422), termed PKMYT1AR (PKMYT1 associated lncRNA), that is induced in NSCLC by Yin Yang 1 (YY1) factor, especially in cancerous cell lines (H358, H1975, H1299, H1650, A549 and SPC-A1) compared to that in normal human bronchial epithelium cell line (BEAS-2B). We show that PKMYT1AR high expression correlates with worse clinical outcome, and knockdown of PKMYT1AR inhibits tumor cell proliferation, migration and xenograft tumor formation abilities. Bioinformatic analysis and a luciferase assay demonstrate that PKMYT1AR directly interacts with miR-485-5p to attenuate the inhibitory role on its downstream oncogenic factor PKMYT1 (the protein kinase, membrane-associated tyrosine/threonine 1) in NSCLC. Furthermore, we uncover that miR-485-5p is downregulated in both cancerous cell lines and peripheral blood serum isolated from NSCLC patients compared to reciprocal control groups. Consistently, forced expression of miR-485-5p inhibits the proliferation and migration abilities of tumor cells. Moreover, we provide evidence showing that PKMYT1AR targeting antisense oligonucleotide (ASO) dramatically inhibit tumor growth in vivo. Mechanistic study shows that PKMYT1AR/ miR-485-5p /PKMYT1 axis promotes cancer stem cells (CSCs) maintenance in NSCLC via inhibiting β-TrCP1 mediated ubiquitin degradation of β-catenin proteins, which in turn causes enhanced tumorigenesis.ConclusionsOur findings reveal the critical role of PKMYT1AR/miR-485-5p /PKMYT1 axis during NSCLC progression, which could be used as novel therapeutic targets in the future.
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影响因子：
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作者：
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通讯作者：
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