Long Non-coding RNA LINC-PINT Suppresses Cell Proliferation and Migration of Melanoma via Recruiting EZH2
Long Non-coding RNA LINC-PINT Suppresses Cell Proliferation and Migration of Melanoma via Recruiting EZH2
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长非编码 RNA LINC-PINT 通过招募 EZH2 抑制黑色素瘤细胞增殖和迁移
DOI：
10.3389/fcell.2019.00350
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发表时间：
2019-12-20
影响因子：
5.5
通讯作者：
Fan, Xianqun
中科院分区：
文献类型：
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作者：
Xu, Yangfan;Wang, Huixue;Fan, Xianqun
长链非编码 RNA (lncRNA) 已被确定为许多人类癌症的关键调节因子。许多lncRNA在癌症中表现出异常表达，其中一些在肿瘤增殖、侵袭和转移中发挥着关键作用。然而，lncRNA 在黑色素瘤进展中的调节功能仍有待阐明。我们利用实时 PCR 方法来确定 LINC-PINT 在黑色素瘤细胞系中的表达。为了评估 LINC-PINT 对黑色素瘤肿瘤发生的影响，我们使用细胞计数试剂盒-8 (CCK8) 和集落形成测定。采用流式细胞术检测LINC-PINT对细胞周期状态的作用。使用 RNA 纯化 (ChIRP) 分离染色质来鉴定 PINT 相互作用蛋白。采用微阵列分析和生物信息学分析来寻找LINC-PINT的潜在靶基因，并使用染色质免疫沉淀分析（ChIP）验证LINC-PINT候选靶基因的状态。 LINC-PINT在抑制黑色素瘤的致瘤性方面发挥作用，这一点通过异种移植模型测定进一步确定。 LINC-PINT 在黑色素瘤组织和细胞系中显着下调。肿瘤细胞中 LINC-PINT 的过度表达导致 A375、Mum2B 和 CRMM1 细胞中肿瘤生长显着减少并抑制迁移。基于体内异种移植模型的结果与 LINC-PINT 阻碍黑色素瘤细胞生长和转移的体外研究结果进一步一致。微阵列分析和生物信息学分析表明CDK1、CCNA2、AURKA和PCNA是LINC-PINT的潜在靶标。总之，LINC-PINT通过将EZH2招募到其靶基因的启动子，导致H3K27三甲基化和靶基因的表观遗传沉默来抑制黑色素瘤的致瘤性。 LINC-PINT 可能作为黑色素瘤的新诊断和治疗靶点。
Long non-coding RNAs (lncRNAs) have been identified as crucial regulators in many human cancers. Many lncRNAs show aberrant expression in cancer, and some of them play critical roles in tumor proliferation, invasion, and metastasis. However, the regulatory functions of lncRNAs in melanoma progression remain to be elucidated. We utilized the Real-time PCR methodology to determine the expression of LINC-PINT in melanoma cell lines. To evaluate the effect of LINC-PINT on tumorigenesis of melanoma, we used Cell Counting Kit-8 (CCK8) and colony formation assay. Flow cytometry assay was used to detect the function of LINC-PINT on cell cycle status. PINT-interacting proteins were identified by chromatin isolation using RNA purification (ChIRP). Microarray assay and bioinformatics analysis were used to find the potential target genes of LINC-PINT and the status of LINC-PINT target gene candidate was verified using chromatin immunoprecipitation assay (ChIP). LINC-PINT plays a role in suppressing the tumorigenicity of melanoma, which was further determined by xenograft model assay. LINC-PINT was significantly downregulated in melanoma tissues and cell lines. The overexpression of LINC-PINT in tumor cells resulted in significant tumor growth reduction and migration inhibition in A375, Mum2B and CRMM1 cells. Results based on the in vivo xenograft model were further consistent with the in vitro findings that LINC-PINT impeded growth and metastasis of melanoma cells. Microarray assay and bioinformatics analysis indicated that CDK1, CCNA2, AURKA, and PCNA were potential targets of LINC-PINT. In conclusion, LINC-PINT inhibits the tumorigenicity of melanoma through recruiting EZH2 to the promoter of its target genes, leading to H3K27 trimethylation and epigenetic silencing of target genes. LINC-PINT may serve as a novel diagnostic and therapeutic target for melanoma.