METTL3 Promotes Growth Of Hormone Sensitive Prostate Cancer By RNA N6-methyladenosine (m6A)

Objective: methyltransferase 3(METTL3)was detected in prostate cancer in vitro and in vivo.The mechanism of METTL3 in the development of hormone-sensitive prostate cancer.Methods: immunohistochemistry was used to detect prostate cancer and benign prostate tissue METTL3.Image J software was used for quantitative analysis.The contents of METTL3 in BPH1 and prostatic cancer cell lines were detected by Western Blot and q PCR in vitro.Sh RNA transfected with lentivirus were used to construct stable prostate cancer cell lines that interfere with METTL3.Western Blot was used to verify the efficiency of knocking down METTL3 in prostate cancer cells.In vitro cell experiments,MTT and cell colony formation experiments were conducted to verify the effect of METTL3 on the proliferation of prostate cancer cells.High-throughput methylation sequencing(m6A RNA-seq)was used to sequence the tissues of five patients with benign prostate tissue and prostate cancer.The differentially expressed genes were compared and the signal pathways were enriched and screened.Western Blot and q PCR were used to verify the expression of enriched differential genes in cell lines.The relationship between METTL3 and m6 A and target genes AKT and c-MYC YAP1 and its potential mechanism were verified by RIP-q PCR.Results: compared with benign prostate tissue,immunohistochemistry showed that METTL3 was highly expressed in prostate cancer tissues,with statistically significant difference(P<0.001).The RNA level and protein level of METTL3 in each prostate cancer cell line were higher than that of BPH1.Down-regulation of METTL3 in two prostate cancer cell lines can significantly reduce the proliferation capacity of prostate cancer cells.METTL3 was down-regulated in prostate cancer cell lines,and the overall RNA m6 A methylation level in prostate cancer cells was decreased.Changes in the abundance of m6 A in hormone-sensitive prostate cancer tissues.Abnormal activation of genes in the Hippo pathway in HSPC.The knockdown of METTL3 will lead to the decrease of m6 a level of m RNA of YAP1 and c-MYC,and the decrease of expression of YAP1,c-MYC and other related genes.Conclusion: METTL3 is abnormally expressed in hormone-sensitive prostate cancer and is significantly increased compared with benign prostate cancer.METTL3 will affect the cell proliferation ability of HSPC cell line,and knocking down METTL3 will weaken the cell proliferation ability.In HSPC,METTL3 mediates the change of m6 A,which further affects the abnormal activation of Hippo-YAP pathway and the abnormal expression of YAP1,c-MYC and other genes.The reduction of METTL3 in prostate cancer cells will significantly reduce the level of YAP1,c-MYC,Cyclin D and other genes,and m6 A.This leads to the decrease of its expression level.