Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences
RBM15/IGF2BP1-mediated m6A methylation of AKR1B10 promotes the progression of cervical squamous cell carcinoma
By Chen Y, Jiang Y, Wang Q and 4 others.
Abstract
Objectives
RNA-binding motif protein 15 (RBM15) is a methyltransferase involved in N6-methyladenosine (m6A) modification and has been shown to be associated with the initiation and progression of various malignancies. This study aims to investigate the biological function and underlying mechanism by which RBM15 promotes the progression of cervical squamous cell carcinoma (CSCC) through in vitro and in vivo experiments.
Methods
The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were used to analyze differences in the mRNA expression levels of RBM15, aldo-keto reductase family 1 member B10 (AKR1B10), and insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) between CSCC and adjacent non-tumor tissues, as well as the associations of RBM15, AKR1B10, and IGF2BP1 expression with overall survival in patients with CSCC. Real-time PCR and Western blotting (WB) were used to determine the expression levels of RBM15, AKR1B10, and IGF2BP1 in CSCC tissues and adjacent normal tissues. Cell Counting Kit-8 (CCK-8), colony formation, 5-ethynyl-2'-deoxyuridine (EdU) staining, wound-healing, and Transwell migration and invasion assays were performed to investigate the effects of RBM15 knockdown on the proliferation, migration, and invasion of CSCC cells. A nude mouse xenograft model was established to evaluate the effect of RBM15 knockdown on tumor growth. Methylated RNA immunoprecipitation (MeRIP) was performed to assess m6A modification of AKR1B10. RNA immunoprecipitation (RIP) and actinomycin D assays were used to analyze the interactions of RBM15 and IGF2BP1 with AKR1B10 mRNA. Small interfering RNAs (siRNAs), including si-IGF2BP1 and si-YTHDF1, were used to knock down IGF2BP1 or YTH N6-methyladenosine RNA-binding protein F1 (YTHDF1), respectively, and their roles in regulating AKR1B10 mRNA stability were analyzed.