Fujita Maika, Kaneda Masahiro
Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
J Vet Med Sci. 2017 Aug 4;79(8):1352-1358. doi: 10.1292/jvms.17-0179. Epub 2017 Jun 24.
DNA methylation is an epigenetic mechanism controlling gene expression without affecting DNA sequences, and aberrant DNA methylation patterns are features of a number of diseases. Notably, epigenetic errors in cancer cells have been intensively studied over the last two decades in humans; however, little is known concerning dogs and cats. To analyze DNA methylation and gene expression changes in feline lymphoma cells, we added the DNA methylation inhibitor 5-aza-2'-deoxycytidine (5-aza) to three cell lines (3281 and FT-1 cells derived from T-cell lymphoma and MS4 cells derived from B-cell lymphoma). Adding 5-aza significantly retarded cell growth in a dose-dependent manner in all cell lines, and there were aberrant gene expression patterns. Transcription factor Sox11 expression in 3281 cells was de-repressed by 5-aza treatment, and subsequent promoter DNA demethylation was analyzed by bisulfite sequencing. Cell cycle analysis suggested that inhibition of cell growth was due to DNA replication arrest, and this supported the result of increased expression of p27kip1 gene which disturbed cells of 3281 and FT-1 entering the S phase. In this study, 5-aza suppressed the growth of feline lymphoma cells, but further experiments with normal lymph cells are necessary to confirm specificity of this drug treatment and to expand it for clinical use.
DNA甲基化是一种表观遗传机制,可在不影响DNA序列的情况下控制基因表达,异常的DNA甲基化模式是许多疾病的特征。值得注意的是,在过去二十年中,人类对癌细胞中的表观遗传错误进行了深入研究;然而,关于犬猫的情况却知之甚少。为了分析猫淋巴瘤细胞中的DNA甲基化和基因表达变化,我们将DNA甲基化抑制剂5-氮杂-2'-脱氧胞苷(5-aza)添加到三种细胞系中(源自T细胞淋巴瘤的3281和FT-1细胞以及源自B细胞淋巴瘤的MS4细胞)。添加5-aza以剂量依赖性方式显著抑制了所有细胞系的细胞生长,并且存在异常的基因表达模式。5-aza处理使3281细胞中的转录因子Sox11表达去抑制,随后通过亚硫酸氢盐测序分析启动子DNA去甲基化。细胞周期分析表明,细胞生长的抑制是由于DNA复制停滞,这支持了p27kip1基因表达增加的结果,该基因干扰了3281和FT-1细胞进入S期。在本研究中,5-aza抑制了猫淋巴瘤细胞的生长,但需要对正常淋巴细胞进行进一步实验,以确认这种药物治疗的特异性并将其扩展用于临床。