NAT10 Promotes Cervical Cancer Cell Malignant Behavior via N4-acetylcytidine Modification of DDR1 mRNA / 中国生物化学与分子生物学报

ABSTRACT

N-acetyltransferase 10 (NAT10) is a key enzyme involved in the acetylation of mRNA, which regulates the expression of target genes and biological functions of various cancers via ac4C (N4-acetylcytidine) acetylation. However, whether NAT10 is involved in regulating the malignant behavior of cervical cancer is rarely reported. This study investigated the effects and specific mechanism of NAT10 on the growth activity, proliferation capacity, migration and invasion capacity of cervical cancer cells. First, we found a highly expression of NAT10 in cervical cancer and was associated with poor patient prognosis by TCGA database analysis. MTT assays and colony formation assays showed that overexpression of NAT10 promoted the growth activity (P<0. 05) and proliferation ability (P<0. 001) of cervical cancer cells; Transwell migration and invasion assays showed that overexpression of NAT10 promoted the migration (P<0. 01) and invasion (P<0. 05) ability of cervical cancer cells; Western blotting showed that overexpression of NAT10 increased the mesothelial cell marker vimentin and resulted in a decrease in the epithelial cell marker E-cadherin. Bioinformatics analysis exhibited that discoidin domain receptor 1 (DDR1) might be a downstream target gene of NAT10. RNA binding protein immunoprecipitation (RIP) assays showed that NAT10 could directly bind to DDR1 mRNA (P<0. 05), Western blotting and RT-qPCR experiments showed that overexpression of NAT10 significantly increased the expression and mRNA levels of DDR1 (P<0. 05). Furthermore, RNA acetylation co-immunoprecipitation experiments showed that overexpression of NAT10 promoted the acetylation level of DDR1 mRNA (P<0. 001), and EGFP reporter assays confirmed that NAT10 recognized the acetylation site of DDR1 mRNA. The results of mRNA half-life experiments showed that NAT10 increased the stability of DDR1 mRNA (P<0. 05). In conclusion, our research confirms that NAT10 promotes the growth activity, and migration and invasion ability of cervical cancer cells and its specific mechanism is to extend the stability of DDR1 by acetylation modification, thereby increasing its expression levels, which might provide new insights into the molecular mechanism of acetylation modification of mRNA on the pathogenesis of cervical cancers.