经鼻给予叶绿醇治疗可改善携带 MTHFR rs1801133 多态性突变型的复发性胶质母细胞瘤患者的生存率。
Intranasal perillyl alcohol therapy improves survival of patients with recurrent glioblastoma harboring mutant variant for MTHFR rs1801133 polymorphism.
机构信息
Instituto de Biologia, Universidade Federal Fluminense, Niteroi, Rio de Janeiro, ZC, 24020-141, Brazil.
Programa de Pós-graduação em Neurologia, Faculdade de Medicina, Universidade Federal Fluminense, Niteroi, Rio de Janeiro, 24020-141, Brazil.
出版信息
BMC Cancer. 2020 Apr 7;20(1):294. doi: 10.1186/s12885-020-06802-8.
BACKGROUND
Polymorphisms in MTHFR gene influence risk and overall survival of patients with brain tumor. Global genomic DNA (gDNA) methylation profile from tumor tissues is replicated in peripheral leukocytes. This study aimed to draw a correlation between rs1801133 MTHFR variants, gDNA methylation and overall survival of patients with recurrent glioblastoma (rGBM) under perillyl alcohol (POH) treatment.
METHODS
gDNA from whole blood was extracted using a commercially available kit (Axygen) and quantified by spectrophotometry. Global gDNA methylation was determined by ELISA and rs1801133 polymorphism by PCR-RFLP. Statistical analysis of gDNA methylation profile and rs1801133 variants included Mann-Whitney, Kruskal-Wallis, Spearman point-biserial correlation tests (SPSS and Graphpad Prism packages; significant results for effect size higher than 0.4). Prognostic value of gDNA methylation and rs1801133 variants considered survival profiles at 25 weeks of POH treatment, having the date of protocol adhesion as starting count and death as the final event.
RESULTS
Most rGBM patients showed global gDNA hypomethylation (median = 31.7%) and a significant, moderate and negative correlation between TT genotype and gDNA hypomethylation (median = 13.35%; rho = - 0.520; p = 0.003) compared to CC variant (median = 32.10%), which was not observed for CT variant (median = 33.34%; rho = - 0.289; p = 0.06). gDNA hypermethylated phenotype (median = 131.90%) exhibited significant, moderate and negative correlations between TT genotype (median = 112.02%) and gDNA hypermethylation levels when compared to CC (median = 132.45%; rho = - 0,450; p = 0.04) or CT (median = 137.80%; rho = - 0.518; p = 0.023) variants. TT variant of rs1801133 significantly decreased gDNA methylation levels for both patient groups, when compared to CC (d values: hypomethylated = 1.189; hypermethylated = 0.979) or CT (d values: hypomethylated = 0.597; hypermethylated = 1.167) variants. Positive prognostic for rGBM patients may be assigned to gDNA hypermethylation for survivors above 25 weeks of treatment (median = 88 weeks); and TT variant of rs1801133 regardless POH treatment length.
CONCLUSION
rGBM patients under POH-based therapy harboring hypermethylated phenotype and TT variant for rs1801133 had longer survival. Intranasal POH therapy mitigates detrimental effects of gDNA hypomethylation and improved survival of patients with rGBM harboring TT mutant variant for MTHFR rs1801133 polymorphism.
TRIAL REGISTRATION
CONEP -9681- 25,000.009267 / 2004. Registered 12th July, 2004.
背景
MTHFR 基因多态性影响脑肿瘤患者的风险和总生存率。肿瘤组织的全基因组 DNA（gDNA）甲基化谱在周围白细胞中复制。本研究旨在探讨在接受香草醇（POH）治疗的复发性胶质母细胞瘤（rGBM）患者中，rs1801133 MTHFR 变体、gDNA 甲基化与总生存率之间的相关性。
方法
使用市售试剂盒（Axygen）从全血中提取 gDNA，并通过分光光度法定量。通过 ELISA 测定全基因组 gDNA 甲基化，通过 PCR-RFLP 测定 rs1801133 多态性。gDNA 甲基化谱和 rs1801133 变体的统计分析包括 Mann-Whitney、Kruskal-Wallis、Spearman 点双序列相关检验（SPSS 和 Graphpad Prism 包；效应大小大于 0.4 时为显著结果）。gDNA 甲基化和 rs1801133 变体的预后价值考虑了 POH 治疗 25 周时的生存情况，以方案依从性日期为起始计数，以死亡为最终事件。
结果
大多数 rGBM 患者表现出全基因组 gDNA 低甲基化（中位数=31.7%），与 CC 变体（中位数=32.10%）相比，TT 基因型与 gDNA 低甲基化之间存在显著、中度和负相关（中位数=13.35%；rho=-0.520；p=0.003），而 CT 变体（中位数=33.34%；rho=-0.289；p=0.06）则没有观察到这种相关性。gDNA 高甲基化表型（中位数=131.90%）与 TT 基因型（中位数=112.02%）和 gDNA 高甲基化水平之间存在显著、中度和负相关，与 CC（中位数=132.45%；rho=-0.450；p=0.04）或 CT（中位数=137.80%；rho=-0.518；p=0.023）变体相比。与 CC（d 值：低甲基化=1.189；高甲基化=0.979）或 CT（d 值：低甲基化=0.597；高甲基化=1.167）变体相比，rs1801133 的 TT 变体显著降低了两组患者的 gDNA 甲基化水平。对于生存时间超过 25 周的 rGBM 患者（中位数=88 周），gDNA 高甲基化可能具有正预后；而无论 POH 治疗时间长短，rs1801133 的 TT 变体都是如此。
结论
接受 POH 为基础的治疗的 rGBM 患者具有高甲基化表型和 rs1801133 的 TT 变体，其生存率更高。鼻内 POH 治疗减轻了 gDNA 低甲基化的不利影响，并改善了携带 rs1801133 MTHFR 突变的 rGBM 患者的生存。
试验注册
CONEP-9681-25,000.009267/2004。2004 年 7 月 12 日注册。
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