Chen Jieqing, Liao Xinhui, Cheng Jianli, Su Ganglin, Yuan Fen, Zhang Zhongfu, Wu Jianting, Mei Hongbing, Tan Wanlong
Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Department of Urology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China.
Front Cell Dev Biol. 2021 Dec 9;9:777349. doi: 10.3389/fcell.2021.777349. eCollection 2021.
Long-chain non-coding RNA (LncRNA) has been found to play an important role in the regulation of the occurrence and progression of renal cell carcinoma (RCC). In this study, we demonstrated that LncRNA NEAT1 expression and m6A methylation level was decreased in RCC tissues. Further, the downregulated expression level of LncRNA NEAT1 was associated with poor prognosis for RCC patients. Then we used CRIPSR/dCas13b-METTL3 to methylate LncRNA NEAT1 in RCC cells. The results showed that the expression level of LncRNA NEAT1 was upregulated after methylated by dCas13b-METTL3 in RCC cells. And the proliferation and migration ability of RCC cells was decreased after methylated LncRNA NEAT1. Finally, we examined the effect of LncRNA NEAT1 hypermethylation on the transcriptome. We found differentially expressed genes in RCC cells were associated with "cGMP-PKG signaling pathway", "Cell adhesion molecules" and "Pathways in cancer". In conclusion, CRISPR/Cas13b-METTL3 targeting LncRNA NEAT1 m6A methylation activates LncRNA NEAT1 expression and provides a new target for treatment of RCC.
长链非编码RNA(LncRNA)已被发现参与肾细胞癌(RCC)发生发展的调控过程。本研究中,我们发现RCC组织中LncRNA NEAT1的表达及m6A甲基化水平降低。此外,LncRNA NEAT1表达下调与RCC患者的不良预后相关。随后,我们利用CRIPSR/dCas13b-METTL3使RCC细胞中的LncRNA NEAT1发生甲基化。结果显示,在RCC细胞中,经dCas13b-METTL3甲基化后,LncRNA NEAT1的表达水平上调。并且,LncRNA NEAT1甲基化后,RCC细胞的增殖和迁移能力降低。最后,我们检测了LncRNA NEAT1高甲基化对转录组的影响。我们发现RCC细胞中差异表达的基因与“cGMP-PKG信号通路”、“细胞黏附分子”及“癌症相关通路”有关。综上所述,靶向LncRNA NEAT1 m6A甲基化的CRISPR/Cas13b-METTL3激活了LncRNA NEAT1的表达,并为RCC的治疗提供了新靶点。