Department of Pathology, Molecular Medicine and Cancer Research Center of Basic Medicine College, Chongqing Medical University, 1 Yixueyuan Road, Yuzhong, Chongqing, 400016, People's Republic of China.
Med Oncol. 2021 Aug 6;38(9):111. doi: 10.1007/s12032-021-01559-x.
Our previous study clarified the carcinogenic properties of arginine-specific mono-ADP-ribosyltransferase 1 (ART1), which results in a critical post-translational modification that changes the structure and function of proteins and is widely involved in important processes. This study provides, for the first time, a comprehensive transcriptomic analysis of colorectal cancer cells with ART1 silencing by Illumina RNA-Seq and related verification experiments. Lentiviral infection was used to construct a CT-26 cell line with stable knockdown of the ART1 gene, and a whole transcriptome sequencing technique was performed to identify differentially expressed genes (DEGs). GO and KEGG classification/enrichment analyses and verification experiments were performed to determine the role of ART1 in the progression of colorectal cancer. A total of 5552 DEGs, GO functions and KEGG pathways with the highest enrichment, various SNPs, and diverse splicing patterns were identified. Importantly, knockdown of ART1 affected the splicing of certain key genes related to tumor cell growth and downregulated the expression of the key gene PTBP1 for alternative splicing. The overall attenuation of the endoplasmic reticulum unfolded protein response (UPR) signaling pathway caused by the inhibition of mono-ADP-ribosylation of GRP78 could disrupt UPR signaling, leading to the occurrence of apoptosis to impede tumorigenesis. ART1, which is clustered in organelles, may promote the development of colorectal cancer by participating in a variety of new mechanisms, including endoplasmic reticulum stress regulation and alternative splicing, and may be a good clinical drug target for targeted therapy of CRC.
我们之前的研究阐明了精氨酸特异性单 ADP-核糖基转移酶 1（ART1）的致癌特性，它导致了蛋白质结构和功能发生关键性的翻译后修饰，广泛涉及重要的过程。本研究首次通过 Illumina RNA-Seq 对 ART1 沉默的结直肠癌细胞进行了全面的转录组分析，并进行了相关验证实验。利用慢病毒感染构建了稳定敲低 ART1 基因的 CT-26 细胞系，采用全转录组测序技术鉴定差异表达基因（DEGs）。进行了 GO 和 KEGG 分类/富集分析和验证实验，以确定 ART1 在结直肠癌进展中的作用。共鉴定出 5552 个差异表达基因、GO 功能和 KEGG 途径，这些基因/途径具有最高的富集程度，以及各种 SNPs 和不同的剪接模式。重要的是，ART1 的敲低影响了与肿瘤细胞生长相关的某些关键基因的剪接，并下调了关键基因 PTBP1 的可变剪接表达。GRP78 的单 ADP-核糖基化抑制导致内质网未折叠蛋白反应（UPR）信号通路的整体减弱，可能破坏 UPR 信号，导致细胞凋亡以阻碍肿瘤发生。定位于细胞器的 ART1 可能通过参与多种新的机制，包括内质网应激调节和可变剪接，促进结直肠癌的发展，并且可能成为 CRC 靶向治疗的一个良好的临床药物靶点。
