Central Laboratory of Yong-Chuan Hospital, Chongqing Medical University, Chongqing, 402160, China.
Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Sci Rep. 2021 Aug 18;11(1):16755. doi: 10.1038/s41598-021-96156-3.
The long-noncoding RNA colorectal neoplasia differentially expressed (CRNDE) gene has been considered to be crucial in tumor malignancy. Although CRNDE is highly expressed in acute myeloid leukemia (AML), its mechanism of action remains unknown. In this study, GEPIA and qRT-PCR were performed to confirm the expression of CRNDE in AML samples and cell lines, respectively. CRNDE shRNA vectors were transfected to explore the biological functions of CRNDE. The cell proliferation was assessed by the CCK8 assay, while apoptosis and cell cycle distribution were measured by flow cytometry and Western blotting. The results showed that CRNDE was overexpressed in both AML samples and cell lines. CRNDE silencing inhibited proliferation and increased apoptotic rate and cell cycle arrest of KG-1a cells. The luciferase reporter assay coupled with RIP assay revealed that CRNDE act as a ceRNA. Rescue assays demonstrated that the effects of CRNDE silencing could be reversed by miR-136-5p inhibitors. In conclusion, our results expound that the CRNDE/miR-136-5p/MCM5 axis modulates cell progression and provide a new regulatory network of CRNDE in KG-1a cells.
长非编码 RNA 结直肠肿瘤差异表达（CRNDE）基因被认为在肿瘤恶性程度中起关键作用。虽然 CRNDE 在急性髓系白血病（AML）中高表达，但作用机制尚不清楚。在这项研究中，通过 GEPIA 和 qRT-PCR 分别证实了 AML 样本和细胞系中 CRNDE 的表达。通过转染 CRNDE shRNA 载体来探索 CRNDE 的生物学功能。通过 CCK8 测定评估细胞增殖，通过流式细胞术和 Western blot 测定评估细胞凋亡和细胞周期分布。结果表明，CRNDE 在 AML 样本和细胞系中均过表达。CRNDE 沉默抑制了 KG-1a 细胞的增殖，并增加了凋亡率和细胞周期阻滞。荧光素酶报告实验和 RIP 实验表明，CRNDE 作为 ceRNA 发挥作用。挽救实验表明，CRNDE 沉默的作用可以被 miR-136-5p 抑制剂逆转。综上所述，我们的研究结果阐述了 CRNDE/miR-136-5p/MCM5 轴调节细胞进展，并为 KG-1a 细胞中 CRNDE 的调控网络提供了新的见解。
