6mA Gene Modification In Placental Tissue Is Involved In The Development Of Preeclampsia

Objective: To analyze the differential expression of DNA N6-methyladenine(6mA)in placenta tissue between preeclampsia(PE)and normal pregnancy and its molecular mechanism,aim to disclosing the effect of 6mA on PE.Methods: The placenta of PE and normal pregnancy were collected and the genomic DNA of placental trophoblasts was extracted.The DNA fragments were transformed into 100-500 bp,and the whole genome DNA was immunized and coprecipitated by Ch IP-seq and other experimental techniques to isolate the genome DNA fragments.Subsequently,the library was constructed by PCR amplification and fragment screening.High throughput sequencing and data analysis of the quality-controlled library were carried out to construct a genome-wide 6mA methylation map of placenta in PE,analyze its biological information,and preliminarily explore the relationship between 6mA and PE.Finally,the expression level of differentially expressed genes was verified by RT-PCR.Results:(1)Reads showed that the number of unique and repetitive reading lengths(115084349)in PE was higher than that in NC(112026034).Peaks analysis showed that the number of peak in placenta of PE(92064)was lower than that of NC(126487).(2)The number of identical genes in PE and NC was 4 1924,the number of specific genes in PE was 45625,and that in NC was 78226.The results of Gene Ontology(GO)analysis in PE indicated that the differentially expressed genes were mainly composed of mitochondria,microtubule bundles,organelle membranes and telomere complexes,which regulated oxidoreductase activity,translation inhibition activity and death domain formation,and affected apoptotic process and oxidative stress response.The results of KEGG analysis showed that the differentially expressed genes in preeclampsia group were mainly involved in pentose,glucuronic acid conversion,antigen processing and presentation,sphingolipid metabolism regulation and Rig-1-like receptor signaling pathway.(3)The results of KEGG analysis were used to select the preeclampsia differential gene(Rig-I-like receptor signaling pathway)for RT-PCR.The results showed that the transcriptional expression level of related genes in the preeclampsia group was lower than that in the normal control group.CONCLUSION: There are differences in the distribution of 6mA gene modification between PE and NC placenta,and different genes participate in many biological functions of cells.The 6mA methylation of related differential genes is related to the inhibition of Rig-I-like receptor signaling pathway expression in placental trophoblasts of preeclampsia,which may be involved in the pathogenesis of preeclampsia.