通过转录组分析表征tfiie调控基因。

Turkish journal of biology = Turk biyoloji dergisi Pub Date : 2024-10-22 eCollection Date: 2024-01-01 DOI:10.55730/1300-0152.2718
Serdar Baysal
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摘要

背景/目的:以往对通用转录因子ⅱE (GTF2E)的研究表明,它与某些类型的疾病,如结肠癌和毛硫营养不良有关,但GTF2E对细胞过程的整体影响仍未广泛表征。本研究旨在研究和表征GTF2E对基因转录水平的影响,并鉴定与GTF2E相关的细胞过程和疾病。材料和方法:用siGTF2E1或非靶向阴性siRNA以30 nM的浓度转染人结直肠癌细胞系HCT116。72h后,收获细胞,准备进一步分析。利用RNA测序技术对siGTF2E1敲除HCT116细胞(n = 3)及其非靶siRNA阴性对照(n = 3)获得的HCT116细胞系进行全转录组分析。采用MTS法检测细胞活力。结果:与对照组相比,GTF2E1敲低时鉴定出166个基因,敲低组表达差异显著,其中上调基因66个,下调基因100个。发现了一个显著丰富的基因本体术语,涉及碳水化合物结合。发现了一个与B细胞慢性淋巴细胞白血病(B- cll)相关的癌基因。确定了5个与结肠癌相关的基因。11个基因参与了动脉粥样硬化的发展。GTF2E1敲低导致细胞活力下降。结论:GTF2E1敲低组出现多个基因表达改变,其中一些基因与动脉粥样硬化、结肠癌、B-CLL的发生有关。这可能揭示了GTF2E的不同调节作用及其与某些疾病的关系。
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Characterization of TFIIE-regulated genes by transcriptome analysis.

Background/aim: Previous studies on general transcription factor II E (GTF2E) showed that it is associated with certain groups of diseases, such as colon cancer and trichothiodystrophy, but the global effect of GTF2E on cellular processes is still not widely characterized. This study aimed to investigate and characterize the effect of GTF2E on the transcription level of genes and identify the cellular processes and diseases associated with GTF2E.

Materials and methods: The human colorectal carcinoma cell line HCT116 used in the study was transfected at a 30 nM concentration with siGTF2E1 or nontarget negative siRNA. After 72 h, cells were harvested and prepared for further analysis. A whole transcriptome analysis was performed on the HCT116 cell line obtained from the siGTF2E1 knockdown of the HCT116 cells (n = 3) and their nontarget negative siRNA controls (n = 3) using RNA sequencing. Cell viability was tested using an MTS assay.

Results: Compared with the control group, 166 genes were identified at the time of the GTF2E1 knockdown and expressed differentially in the knockdown group, including 66 upregulated genes and 100 downregulated genes. One significantly enriched Gene Ontology term was identified, involving carbohydrate binding. One oncogene related to B cell chronic lymphocytic leukemia (B-CLL) was identified. Five genes associated with colon carcinoma were determined. Eleven genes involved in the development of atherosclerosis were identified. GTF2E1 knockdown caused a decrease in cell viability.

Conclusion: The GTF2E1 knockdown group exhibited an altered expression of multiple genes, some of which are related to the development of atherosclerosis, colon carcinoma, and B-CLL. This might shed light on the different regulatory effects of GTF2E and its association with certain diseases.

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