Asa Tasnima Alam, Singh Chabungbam Dhurbachandra, Singh Thokchom Simander, Salahi Saleh, Alom Kazi Morshed, Seo Young Jun
Department of Chemistry, Jeonbuk National University, Jeonju 54896, South Korea.
Department of Chemistry, Jeonbuk National University, Jeonju 54896, South Korea.
Bioorg Chem. 2025 Apr;157:108328. doi: 10.1016/j.bioorg.2025.108328. Epub 2025 Mar 1.
In this study, we employed imidazole-activated natural or modified guanosine derivatives to extend the 3' ends of mRNA using a nonenzymatic method beyond 30 poly-A tails. We evaluated their impact on the translation activity in cell studies using three genes: GFP, Luciferase, and Apoptin. The assessments were conducted through cell imaging, fluorescence, luminescence, western blot analysis, and RT-qPCR to evaluate varying apoptosis-mediated EZH2 expression in cancer epigenetics, among the compounds tested GMP-2-amino-IM, 2'O-Me-2-amino-IM, and N7-(2-MePy)-GMP-IM. The sugar-modified 2'O-Me-GMP-2-amino-IM demonstrated the most favorable results as mRNAs treated with this compound exhibited higher expression levels with promising mRNA stability relative to the control mRNA (without any extension) and other tested compounds. Subsequently, we transfected cancer cells with nonenzymatically modified apoptin mRNAs by utilizing the three imidazole-activated guanosine derivatives compounds and monitored the induced apoptosis. These findings suggest that 2'O-Me-2-amino-IM-modified apoptin mRNA could serve as a promising tool for cancer therapy by inducing apoptosis while selectively modulating EZH2 expression, a key regulator in oncogene suppression.
在本研究中,我们使用咪唑活化的天然或修饰的鸟苷衍生物,通过非酶促方法将mRNA的3'末端延伸至超过30个聚腺苷酸尾巴。我们使用绿色荧光蛋白(GFP)、荧光素酶和凋亡素三个基因,在细胞研究中评估了它们对翻译活性的影响。通过细胞成像、荧光、发光、蛋白质免疫印迹分析和逆转录定量聚合酶链反应(RT-qPCR)进行评估,以评估癌症表观遗传学中不同凋亡介导的EZH2表达,在所测试的化合物中包括GMP-2-氨基-IM、2'-O-甲基-2-氨基-IM和N7-(2-甲基吡啶)-GMP-IM。糖修饰的2'-O-甲基-GMP-2-氨基-IM显示出最有利的结果,因为用该化合物处理的mRNA相对于对照mRNA(无任何延伸)和其他测试化合物表现出更高的表达水平以及良好的mRNA稳定性。随后,我们利用三种咪唑活化的鸟苷衍生物化合物对癌细胞进行非酶促修饰的凋亡素mRNA转染,并监测诱导的凋亡。这些发现表明,2'-O-甲基-2-氨基-IM修饰的凋亡素mRNA可以通过诱导凋亡同时选择性调节EZH2表达(癌基因抑制中的关键调节因子),成为一种有前景的癌症治疗工具。