Varol Nuray, Keles İbrahim, Yildiz Handan, Karaosmanoglu Cem, Karalar Mustafa, Zengin Kursad, Sarici Hasmet, Tokyol Cigdem
Department of Medical Biology and Genetics, Faculty of Medicine, Gazi Univesity, Ankara, Turkey.
Department of Urology, Faculty of Medicine, Afyonkarahisar Health Sciences University, Afyonkarahisar, Turkey.
Gene. 2023 May 25;866:147352. doi: 10.1016/j.gene.2023.147352. Epub 2023 Mar 8.
Recently, aberrant DNA methylation of the HIST1H4F gene (encodes Histone 4 protein) has been shown in many types of cancer, which may serve as a promising biomarker for early cancer diagnosis. However, the correlation between DNA methylation of the HIST1H4F gene and its role in gene expression is unclear in bladder cancer. Therefore, the first objective of this study is to explore the DNA methylation pattern of the HIST1H4F gene and then further elucidate its effects on HIST1H4F mRNA expression in bladder cancer. To this end, the methylation pattern of the HIST1H4F gene was analyzed by pyrosequencing and the effects of the methylation profiles of this gene on HIST1H4F mRNA expression in bladder cancer were examined by qRT-PCR. Sequencing analysis revealed significantly higher methylation frequencies of the HIST1H4F gene in bladder tumor samples compared to normal samples (p < 0,0001). However, when we evaluated the correlations between hypermethylation of HIST1H4F and the clinicopathological parameters (tumor stage, tumor grade, lymph node metastasis, muscle-invasion), no significant difference was found between the groups (p > 0.05). In addition, we examined the role of hypermethylation of the HIST1H4F gene on HIST1H4F mRNA expression. We found that hypermethylation of HIST1H4F in the exon have no effect HIST1H4F mRNA expression in bladder cancer (p > 0.05). We also confirmed our finding in cultured T24 cell line which HIST1H4F gene is hypermethylated. Our results suggest that hypermethylation of the HIST1H4F seems to be a promising early diagnostic biomarker in bladder cancer patients. However, further studies are needed to determine the role of HIST1H4F hypermethylation in tumorigenesis.
最近，HIST1H4F基因（编码组蛋白4蛋白）的异常DNA甲基化在多种癌症中均有发现，这可能是一种很有前景的早期癌症诊断生物标志物。然而，在膀胱癌中，HIST1H4F基因的DNA甲基化与其在基因表达中的作用之间的相关性尚不清楚。因此，本研究的首要目标是探索HIST1H4F基因的DNA甲基化模式，进而阐明其对膀胱癌中HIST1H4F mRNA表达的影响。为此，通过焦磷酸测序分析了HIST1H4F基因的甲基化模式，并通过qRT-PCR检测了该基因甲基化谱对膀胱癌中HIST1H4F mRNA表达的影响。测序分析显示，与正常样本相比，膀胱癌样本中HIST1H4F基因的甲基化频率显著更高（p < 0.0001）。然而，当我们评估HIST1H4F基因高甲基化与临床病理参数（肿瘤分期、肿瘤分级、淋巴结转移、肌肉浸润）之间的相关性时，各组之间未发现显著差异（p > 0.05）。此外，我们研究了HIST1H4F基因高甲基化对HIST1H4F mRNA表达的作用。我们发现，外显子中HIST1H4F基因的高甲基化对膀胱癌中HIST1H4F mRNA表达没有影响（p > 0.05）。我们还在HIST1H4F基因高甲基化的培养T24细胞系中证实了这一发现。我们的结果表明，HIST1H4F基因的高甲基化似乎是膀胱癌患者有前景的早期诊断生物标志物。然而，需要进一步研究来确定HIST1H4F基因高甲基化在肿瘤发生中的作用。
