AI Article Synopsis
- The study investigates the role of lncRNA CARMN as a tumor suppressor in cervical cancer (CC) and explores how mA modification and miRNAs, specifically miR-21-5p, affect its expression.
- Using methods like MeRIP-seq and RT-qPCR, researchers found that CARMN has higher mA modification in tumor tissues, and that the mA reader YTHDF2 facilitates its degradation in CC cells.
- Additionally, miR-21-5p was identified as a direct regulator of CARMN, negatively impacting its levels, which suggests that both mA modification and miR-21-5p are critical in the progression of cervical cancer.
Article Abstract
Purpose: The abnormal regulation of lncRNA CARMN has been proved to be a tumor suppressor gene of cervical cancer (CC). However, its role in CC is still elusive. The regulation of CARMN post-transcriptional level by mA modification and miRNA has not been studied. This study aims to analyze the molecular mechanism of mA modification and miRNA on the abnormal expression of CARMN in CC cells, so as to provide a new theoretical basis for the diagnosis and treatment of CC.
Methods: MeRIP-seq was used to identify the differential mA-modified genes between tumor and normal cervical tissues. RT-qPCR assay was used to detect gene expression levels in tissues or cells. The mA modification sites of CARMN was predicted by bioinformatics, and the modification of mA and its regulatory effect on CARMN were analyzed by MeRIP-qPCR, Actinomycin D assay and RIP assay. RIP-microarray combined with bioinformatics methods to screen miRNAs that may target CARMN. The regulation mechanism between miRNA and CARMN was verified by RT-qPCR, nucleo-plasmic separation assay, mRNA stability assay, dual-luciferase reporter assay, and in vivo experiments.
Results: MeRIP-seq found that CARMN is a significant different gene in the abundance of mA in CC, and the modification level of mA in CC tissues was higher than that in normal cervical tissues. Further, this study verified that mA reader YTHDF2 could recognize mA-modified CARMN and promote its degradation in CC cells. miR-21-5p was proved to be the downstream target gene of CARMN, and miR-21-5p could negatively regulate the expression of CARMN. Further experiments showed that miR-21-5p could directly bind to CARMN and lead to the degradation of CARMN. The in vivo experimental results indicated that the level of miR-21-5p in the overexpressed CARMN group was significantly lower than that in the control group.
Conclusion: mA modification and miR-21-5p play important roles in promoting the occurrence and development of tumors by regulating CARMN, provide new potential targets for the treatment of CC.
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