5-Aza-CdR promotes partial MGMT demethylation and modifies expression of different genes in oral squamous cell carcinoma
5-Aza-CdR promotes partial MGMT demethylation and modifies expression of different genes in oral squamous cell carcinoma
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DOI：
10.1016/j.oooo.2019.01.006
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发表时间：
2019-05-01
影响因子：
2.9
通讯作者：
de Souza, Ana Paula
中科院分区：
文献类型：
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作者：
do Amaral, Guilherme C. L. S.;Planello, Aline C.;de Souza, Ana Paula
Objective.口腔鳞状细胞癌（OSCC）的治疗策略因诊断阶段而异。手术和放疗是I期患者局部病变的选择，而化疗是转移性OSCC的主要治疗方法。然而，侵袭性肿瘤可能复发，经常导致死亡。为了解决这个问题，使用改变表观遗传特征的药物的新治疗方案已经成为控制肿瘤生长和转移的替代方案。因此，本研究的目的是研究去甲基化药物5-aza-CdR在SCC 9 OSCC细胞中的作用。用浓度为0.3 μ M和2 μ M的5-Aza-CdR处理SCC 9细胞24小时和48小时。通过使用甲基化特异性高分辨率熔解技术研究MGMT、BRCA 1、APC、c-MYC和hTERT基因的DNA甲基化。采用真实的时间-聚合酶链反应和定量聚合酶链反应分析基因表达。5-Aza-CdR促进了MGMT的去甲基化，并改变了所有分析基因的转录。奇怪的是，浓度为0.3 μ M的5-aza-CdR在SCC 9细胞中比2 mM更有效。我们观察到5-aza-CdR导致MGMT去甲基化，上调3个重要肿瘤抑制基因的转录，并促进c-Myc的下调。
Objective. Treatment strategies for oral squamous cell carcinoma (OSCC) vary, depending on the stage of diagnosis. Surgery and radiotherapy are options for localized lesions for stage I patients, whereas chemotherapy is the main treatment for metastatic OSCC. However, aggressive tumors can relapse, frequently causing death. In an attempt to address this, novel treatment protocols using drugs that alter the epigenetic profile have emerged as an alternative to control tumor growth and metastasis. Therefore, the objective in this study was to investigate the effect of the demethylating drug 5-aza-CdR in SCC9 OSCC cells.Study Design. SCC9 cells were treated with 5-Aza-CdR at concentrations of 0.3 mu M and 2 mu M for 24 hours and 48 hours. DNA methylation of the MGMT, BRCA1, APC, c-MYC, and hTERT genes were investigated by using the methylation-specific high-resolution melting technique. Real time-polymerase chain reaction and quantitative polymerase chain reaction were performed to analyze gene expression.Results. 5-Aza-CdR promoted demethylation of MGMT and modified the transcription of all analyzed genes. Curiously, 5-aza-CdR at the concentration of 0.3 mu M was more efficient than 2 mM in SCC9 cells.Conclusions. We observed that 5-aza-CdR led to MGMT demethylation, upregulated the transcription of 3 important tumor suppressor genes, and promoted the downregulation of c-Myc.