Transcription Silencing and CpGs Hypermethylation as Therapeutic Gene Editing in Clinical Colorectal Adenocarcinoma Repression.

Miqdam M Obaid Al-Jumaili
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引用次数: 0

Abstract

Background/aims: Colorectal cancer is the most common cancer in oncopathology, with an increasing incidence among the elderly during the last decade. Various genetic and environmental factors play important roles in the emergence of colorectal adenocarcinoma. Non-coding RNAs, approximately 20-22 nucleotides, are transcribed irregularly in many cancer cells and play a critical role in many metabolic pathways in clinical cancer cases. DNA methylation is a critical epigenetic alteration that controls gene expression. In the current study, transcriptional silencing and CpG hypermethylation were developed as a therapeutic gene editing strategy for the clinical repression of colorectal adenocarcinoma.

Methods: A human colorectal adenocarcinoma cell line (Caco2) and a normal lung fibroblast cell line (Wi38) were utilized as the paradigms in this research to examine the effect of mir155 molecule transfection and CpGs-island (CGI) methylation. Cell counting was achieved using six-well and 24-well plates before transfection using a hemocytometer. The two cell lines were transfected with the mir155 agomir and antagomir molecules. The transfection efficiency, cell viability, cell IC50, and target gene expression were measured, and CGIs-methylation was achieved by bisulfate conversion.

Results: The outcomes revealed the downregulation of oncogenes (AKT1 and VCAM1 genes as cancer-associated genes) and the upregulation of tumor suppressor genes (TSGs, Tp53 and KEAP1). In addition, CpG-islands methylation showed significant blocking of the oncogene promoter regions, and the switch on of TSG promoter regions was continuous.

Conclusions: miRNA-CGI-methylation led to the regression of Caco2 cell proliferation, suggesting the potential use of RNA silencing and DNA methylation in targeted gene therapy for colorectal cancer.

转录沉默和 CpGs 高甲基化作为治疗性基因编辑在临床大肠腺癌抑制中的应用。
背景/目的:结直肠癌是肿瘤病理学中最常见的癌症,近十年来老年人的发病率不断上升。各种遗传和环境因素在结直肠腺癌的发生中起着重要作用。非编码 RNA(约 20-22 个核苷酸)在许多癌细胞中不规则转录,在临床癌症病例的许多代谢途径中发挥关键作用。DNA 甲基化是控制基因表达的重要表观遗传学改变。在目前的研究中,转录沉默和 CpG 高甲基化被开发为一种治疗性基因编辑策略,用于临床抑制结直肠腺癌:方法:以人结直肠腺癌细胞系(Caco2)和正常肺成纤维细胞系(Wi38)为研究范例,考察mir155分子转染和CpGs岛(CGI)甲基化的效果。转染前使用血细胞计数器对 6 孔板和 24 孔板进行细胞计数。用 mir155 agomir 和 antagomir 分子转染两种细胞系。转染效率、细胞存活率、细胞 IC50 和靶基因表达量进行了测定,并通过硫酸氢盐转化实现了 CGIs 甲基化:结果显示,癌基因(AKT1 和 VCAM1 癌相关基因)下调,抑癌基因(TSGs、Tp53 和 KEAP1)上调。结论:miRNA-CGI-甲基化可抑制 Caco2 细胞的增殖,表明 RNA 沉默和 DNA 甲基化可用于结直肠癌的靶向基因治疗。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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