PRMT5 Coordinates With Snail And The NuRD(MTA1) Complex To Promote Metastasis Of Cervical Cancer By Epigenetic Regulation

Objectives: The aim of this study is to gain a deeper understanding of the role of PRMT5 in tumorigenesis and metastasis of cervical cancer by exploring the interaction mechanisms and functions of PRMT5,Snail and Nu RD(MTA1)complex.We want to speculate the possibility of PRMT5 as a new target for tumor detection and treatment.These are specifically divided as following:1.Identifying proteins that are associated with PRMT5 and explore the interaction mechanisms and functions of these proteins.2.To investigate the roles and manners of PRMT5/Snail/Nu RD(MTA1)complex in transcriptional regulation.3.To gain a better view of PRMT5 in the invasion and metastasis of cervical cancer and other tumors.Methods:1.Identifying proteins that are associated with PRMT5 by immunoaffinity purification combined with silver-stained mass spectrometry analysis.Co-IP and FPLC assays are performed to verify the results of mass spectrometry analysis.GST pulldown assays are implemented to further explore the molecular basis for the interaction between these proteins.2.Ch IP assays are combined with Ch IP-seq sequencing analysis to find out target genes that are regulated by PRMT5/Snail/Nu RD(MTA1)complex.The q Ch IP assays are performed to verify the target genes regulated by this complex and to explore the ways to transcriptionally regulates the target genes.3.The cervical cancer cell lines are used for in vitro experiments to detect the expression of EMT markers,wound healing efficiency and transwell ability;the NOD-SCID mice are used to construct animal models to detect the tumorigenesis and metastasis of cervical cancer cells in vivo.Results:1.PRMT5 can specifically interact with Snail and Nu RD(MTA1)to form a complex with epigenetic regulatory functions.2.The PRMT5/Snail/Nu RD(MTA1)complex can participate in transcriptional regulation to suppress the expression of target genes.The members of this complex play synergistically during the transcriptional regulation of target genes.Snail as a transcription factor could identify then combinate with the promoter region of the target genes;PRMT5 and Nu RD(MTA1)are recruited subsequently to catalyze the symmetric dimethylation and deacetylation of histones,resulting in more compacted chromatin and suppression of the target genes E-cadherin,?-catenin and GATA3.3.PRMT5 can promote invasion and metastasis of cervical cancer cells both in vitro and in vivo.In addition,the spheroid forming assays are performed to prove that PRMT5 level is positively correlated with the stemness of cervical cancer cells,which further confirming that PRMT5 plays an important role in the invasion and metastasis ability of cervical cancer cells.4.Immunohistochemical analysis indicate that the expression of PRMT5 is upregulated in cervical cancer tissues and positively correlated with pathological grade.We also find that PRMT5 is highly expressed in various cancer tissues.Conclusion: To sum up,our data has identified many proteins that are associated with PRMT5,and proved that PRMT5/Snail/Nu RD(MTA1)complex participate in the regulation of EMT and metastasis of tumor cells by inhibiting the expression of E-cadherin,?-catenin and GATA3,which promote tumorigenesis and metastasis of cervical cancer or other cancers.We have proved that PRMT5 is highly expressed in cervical cancer and multiple tumor tissues,suggesting that PRMT5 may be a potential tumor marker and a new target for the diagnosis and treatment of cervical cancer or other tumors.