Molecular Mechanism Of YTHDC2 In Promoting Prostate Cancer Progression

The m6 A is a widespread base modification modifier on m RNAs.The m6 A is regulated by the methyltransferase(METTL3,METTL14,and WTAP),demethylases(FTO and ALKBH5)and the corresponding readers(YTHDF1/2/3,YTHDC1/2).Studies have reported that the overall modification levels of m6 A and the expression levels of proteins such as Writer/Reader/Eraser are often dysregulated in many types of cancer and are critical for the tumor development,progression,metastasis,drug resistance,and recurrence of cancers.The function of m6 A is recognized by m6 A binding proteins,i.e.YTH family members(YTHDF1,YTHDF2,YTHDF3,YTHDC1,YTHDC2)and the HNRNP family members of nuclear heterogeneity proteins(HNRNPA2B1 and HNRNPC).In recent years,YTHDC2 has become a hot topic in this research area due to the in-depth study of m6 A modification.As an important m6 A binding protein,YTHDC2 regulates the translation of target genes by specifically recognizing the m6 A sites.YTHDC2 is highly expressed in mouse testis,and knocked out of YTHDC2 in mice shows a 37%reduction in germ cell development and sterility.It has also been suggested that YTHDC2 may be associated with the development of malignant tumors.YTHDC2 may be a potential susceptibility gene for pancreatic cancer and plays an important role in the development of liver cancer,but the relationship between YTHDC2 and prostate cancer has rarely been investigated.The study provides an in-depth study of the interrelatedness of the RNA reading protein YTHDC2 and PCa,elucidates the mechanism of action of YTHDC2,and explores the effect of YTHDC2 on PCa development and its mechanism at the molecular,cellular,tissue,and animal levels.The main findings are as follows:1.YTHDC2 is positively associated with clinical features of prostate cancer in public databases.In the study,we investigated whether YTHDC2 is relevant to the diagnosis and prognosis of prostate cancer.We analyzed the expression of YTHDC2 in prostate cancer patients and its relationship with prognosis in a comprehensive manner in several public databases,including the GEO and TCGA databases.Our results showed that YTHDC2 was highly expressed in PCa tissues,and YTHDC2 expression was positively correlated with T-stage,lymph node metastasis,and Gleason score.These results suggest that the m RNA expression levels of YTHDC2 are positively correlated with the malignant biology of prostate cancer and may act as an independent prognostic factor.GSEA analysis demonstrates that YTHDC2 is involved in cancer-related pathways.The above-mentioned findings suggest that YTHDC2 may be a potential biomarker for tumor development.2.The immunohistochemical scores of YTHDC2 in clinical samples are significantly and positively correlated with clinical malignant features of PCaThe immunohistochemical analysis was applied to the tissue microarray of prostate cancer samples and our prostate samples.YTHDC2 protein was found to be highly expressed in PCa tissues with statistically significant differences(p<0.05),and higher expression of YTHDC2 was significantly or highly significantly correlated with patient's T-stage,clinical-stage,PSA level,and Gleason score(p<0.05),but not with age.3.Interference with YTHDC2 protein significantly inhibits the proliferation,migration and invasion of PC3 cellsWhen si RNA was applied to interfere with YTHDC2,it was found that si YTHDC2 was able to inhibit the proliferation,migration,and invasion ability of DU145 cells,promote cell apoptosis.And conversely,overexpression of YTHDC2 was able to promote the proliferation,migration,and invasion ability of PC3 cells,inhibit cell apoptosis.4.Generation of the in vivo xenograft modelInterfering with YTHDC2 in DU145 cells significantly suppressed the growth and metastasis of tumor xenografts growth in vivo.5.RNA sequencing combined with m6 A sequencing to screen potential downstream target genesCombined RNA-Seq and Me RIP to search for YTHDC2 downstream target genes and pathways.In summary,through bioinformatic analysis,the study found that YTHDC2 was highly expressed in PCa and significantly correlated with clinical traits,and was associated with progression-free survival in prostate cancer patients,indicating that the m RNA expression level of YTHDC2 was positively correlated with the malignant biological properties of prostate cancer,and it could be an independent prognostic factor.The GSEA analysis suggested that YTHDC2 was involved in cancer-related pathways.These results suggested that YTHDC2 may be a potential biomarker for tumor development.Interference with YTHDC2 protein significantly inhibited cell proliferation,migration,and invasion of DU145 cells.And conversely,overexpression of YTHDC2 protein significantly promoted cell proliferation,migration,and invasion of PC3 cells.In vitro and in vivo experiments revealed that interference with YTHDC2 in DU145 cells inhibited tumor growth and metastasis.To clarify the molecular mechanism of YTHDC2 affecting the proliferation of prostate cancer cells,it is of great theoretical value and application to establish new ways and methods for the treatment of prostate cancer.