Immunogenetics Research Center, Department of Tissue Engineering and Applied Cell Sciences, Faculty of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran.
Molecular and Cell Biology Research Center, Mazandaran University of Medical Sciences, Sari, Iran.
Mol Biol Rep. 2024 Jan 9;51(1):96. doi: 10.1007/s11033-023-08934-z.
Rn7SK, a highly conserved small nuclear non-coding RNA, controls Polymerase II transcription machinery by activating of the Positive Transcriptional Elongation Factor b (P-TEFb). Apart from its role in transcriptional regulation, the potential functions of Rn7SK in cell apoptosis are poorly understood. In a previous study, we demonstrated that overexpression of 7SK induces apoptosis in HEK cells. However, it remains unclear whether 7SK-mediated apoptosis induction is exerted through the intrinsic or extrinsic pathways.
Rn7SK was overexpressed in HEK 293T cell line using Lipofectamine 2000 reagent to investigate its potential apoptotic functions. The overexpression of Rn7SK resulted in reduced cell viability through the induction of apoptosis, as evidenced by MTT assay and Annexin V/PI staining. Concurrently, alterations in the expression levels of key apoptosis-related genes were observed, as determined by quantitative RT-PCR. Furthermore, Rn7SK overexpression led to a decrease in cell proliferation, as assessed by colony formation assay and growth curve analysis. This reduction was associated with downregulated expression of key proliferative-related genes. Additionally, the migration and invasion capabilities of cells were significantly inhibited upon upregulation of Rn7SK, as demonstrated by transwell assays.
This study suggests the apoptotic role of 7SK through both intrinsic and extrinsic pathways, necessitating further investigation into its underlying mechanisms.
Rn7SK 是一种高度保守的小核非编码 RNA，通过激活正转录延伸因子 b（P-TEFb）来控制 Polymerase II 转录机制。除了在转录调控中的作用外，Rn7SK 在细胞凋亡中的潜在功能还知之甚少。在之前的研究中，我们证明了 7SK 的过表达会诱导 HEK 细胞凋亡。然而，7SK 介导的凋亡诱导是否通过内在或外在途径发挥作用尚不清楚。
使用 Lipofectamine 2000 试剂在 HEK 293T 细胞系中过表达 Rn7SK，以研究其潜在的凋亡功能。MTT 检测和 Annexin V/PI 染色证实，7SK 的过表达通过诱导细胞凋亡导致细胞活力降低。此外，通过定量 RT-PCR 检测到关键凋亡相关基因的表达水平发生变化。此外，集落形成实验和生长曲线分析表明，Rn7SK 的过表达导致细胞增殖减少，这与关键增殖相关基因的下调表达有关。此外，通过 Transwell 实验证实，细胞的迁移和侵袭能力在 Rn7SK 的上调后显著受到抑制。
本研究表明 7SK 通过内在和外在途径发挥凋亡作用，需要进一步研究其潜在机制。
