5-Aza-dC suppresses melanoma progression by inhibiting GAS5 hypermethylation
5-Aza-dC suppresses melanoma progression by inhibiting GAS5 hypermethylation
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DOI：
10.3892/or.2022.8334
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发表时间：
2022-05
期刊：
影响因子：
4.2
通讯作者：
Yangjie Zhang;R. Xie;Jie Jiang;Li Zhai;Congcong Yang;J. Zhang;Xi Wang;Dongshao Chen;Huatao Niu;Long Chen
中科院分区：
文献类型：
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作者：
Yangjie Zhang;R. Xie;Jie Jiang;Li Zhai;Congcong Yang;J. Zhang;Xi Wang;Dongshao Chen;Huatao Niu;Long Chen
对黑色素瘤发病机制的深入研究表明，表观遗传修饰，特别是DNA甲基化，是黑色素瘤发生和发展的普遍固有特征。本研究对肿瘤抑制基因生长抑制特异性转录本5（GAS 5）的分析表明，其在黑色素瘤中表达下调，且表达水平与其甲基化修饰水平呈一定的负相关。甲基化特异性聚合酶链反应分析表明，在黑色素瘤组织和细胞中，GAS 5的启动子区存在可检测到的CpG岛，而在正常组织中未检测到GAS 5的甲基化修饰。用5-氮杂-2 ′-脱氧胞苷（5-Aza-dC）处理黑色素瘤细胞后，GAS 5甲基化显著逆转。黑色素瘤细胞增殖分析显示，5-Aza-dC以时间依赖性方式抑制A375和SK-MEL-110细胞增殖。进一步的细胞凋亡分析表明，5-Aza-dC显著增加了两种细胞系的凋亡水平。此外，黑色素瘤细胞的迁移分析显示，5-Aza-dC显着降低细胞迁移。此外，5-Aza-dC显著降低了两种细胞系的侵袭能力。当GAS 5表达被沉默时，5-Aza-dC对细胞增殖、凋亡、侵袭和迁移的影响不明显。此外，皮下注射A375细胞在裸鼠中成功地导致异种移植瘤形成。然而，腹腔注射5-Aza-dC后，异种移植瘤的体积和重量以及Ki-67表达显著减少，caspase-3活性和GAS 5表达增强; GAS 5沉默后，5-Aza-dC的抗肿瘤作用被显著阻断。总之，本研究表明5-Aza-dC抑制黑色素瘤的生长，其作用可能与GAS 5的甲基化修饰有关。
The in-depth study of melanoma pathogenesis has revealed that epigenetic modifications, particularly DNA methylation, is a universal inherent feature of the development and progression of melanoma. In the present study, the analysis of the tumor suppressor gene growth arrest-specific transcript 5 (GAS5) demonstrated that its expression was downregulated in melanoma, and its expression level had a certain negative association with its methylation modification level. The promoter of GAS5 presented with detectable CpG islands, and methylation-specific polymerase chain reaction analysis demonstrated that GAS5 was actually modified by methylation in melanoma tissues and cells; however, no methylation modification of GAS5 was detected in normal tissues. Following the treatment of melanoma cells with 5-Aza-2′-deoxycytidine (5-Aza-dC), GAS5 methylation was significantly reversed. The analysis of melanoma cell proliferation revealed that 5-Aza-dC inhibited A375 and SK-MEL-110 cell proliferation in a time-dependent manner. Further analysis of apoptosis demonstrated that 5-Aza-dC significantly increased the apoptosis level of the two cell lines. Moreover, migration analysis of melanoma cells revealed that 5-Aza-dC significantly reduced cell migration. Furthermore, 5-Aza-dC significantly decreased the invasive ability of the two cell lines. However, when the expression of GAS5 was silenced, the effects of 5-Aza-dC on cell proliferation, apoptosis, invasion and migration were not significant. Furthermore, the subcutaneous injection of A375 cells in nude mice successfully resulted in xenograft tumor formation. However, following an intraperitoneal injection of 5-Aza-dC, the volume and weight of xenograft tumors and Ki-67 expression were significantly reduced, and caspase-3 activity and GAS5 expression were enhanced; following the silencing of GAS5, the antitumor effect of 5-Aza-dC was significantly blocked. On the whole, the present study demonstrates that 5-Aza-dC inhibits the growth of melanoma, and its function may be related to the methylation modification of GAS5.