CCDC152 Regulates The Molecular Mechanism Of GPCR Signaling In Pancreatic Cancer Cells

CCDC152(Coiled-coil Domain Containing Protein 152)belongs to the CCDC family.CCDC152 gene is conserved in mammals,but its function is unknown.Bioinformatical prediction showed CCDC 152 tertiary structure is highly similar to that of GCN4 in yeast.The coiled-coil domains have important biological functions in vivo.It often acts as a transcription factor to regulate gene expression.GCN4 lies in yeast nucleus and contains bZIP structure.This structure binds to the corresponding promoter DNA sequences(TGAG/CTCA).GCN4 of yeast usually binds to the specific DNA sequences in dimer manner.Apoptosis and proliferation are essential vital events movement to livings.Apoptosis is a complicated process,which it involves in several apopticsis genes expressions.A large amount of experiments(transcriptiomic results in this lab)suggests CCDC152 may be is involved in apoptosis.Immunohistochemical analysis showed that CCDC 152 was highly expressed in human pancreatic cancer cells(Panc-1 cells).Note that transcription sequencing results for a class of genes CCDC152-overexpression has an largest effect on the GPCR and the downstream signaling systems.According to the basis to explore CCDC 152 in Panc-1 cells the Molecular mechanisms of CCDC 152 and GPCR signaling systems,this experiment adopts immune chemistry,Western blotting,cellular co-localization experiments demonstrated that expression of CCDC 152,which containing coiled coil domain protein family member,localization in cells,with inhibition of GPCR signaling system,Rho/ROCK signal system,Hippo/YAP signal system in Panc-1 cells,and CCDC 152 is a negative regulator of the hub,and can inhibit cell apoptosis and promote cell proliferation.The research main results are as follows:1.Through the extraction of Nuclear/Cytoplasm protein,co-localization experiments proved that CCDC 152 and GCN4 play a role in the nucleus to form two dimers.2.1n Panc-1 cells,GnRH induced CCDC 152 expression through Gai pathway;CCDC 152-overexpression and GnRH with different concentration treated,MTT experiments showed that the cell resistance increases,and induced by CCDC 152 the drug resistance and LY294002,Sulindac,Gefitinib,VPA inhibition signal transduction has a relationship,so CCDC152 can inhibited the GPCR signal transduction induced by GnRH;3.CCDC 152-overexpression can downregulation of the phosphorylation level of the substrate downstream of the Rho/ROCK signaling system and inhibited cell adhesion formation,thus showed that CCDC 152 inhibited the Rho/ROCK signaling system.4.CCDC 152-overexpression can downregulate CTGF protein and p-YAP protein in the Hippo/YAP signaling system,thus proved that the inhibition effect of CCDC 152 on the Hippo/YAP signaling system;Wnt signal and Hippo/YAP signal system is closely linked,CCDC152-overexpression can down regulate the expression of DKK1 protein,which is a negative regulatory factors in Wnt signaling system,thus proved that CCDC 152 has a promoting effect on the Wnt signaling system.5.CCDC 152 is located in the nucleus,As203 or MG-132 stimulates Panc-1 cells,thus proved that CCDC 152 degradation occurs in the nucleus,histone experiment proved that CCDC 152 is involved in the expression of H3 histone modifications associated proteins,Westernblot showed that overexpression of CCDC 152 has an inhibitory effect on PML.6.Cell migration experiments showed that overexpression of CCDC 152 could inhibit cell migration.Conclusion:CCDC 152 expression is regulated by GPCR signaling.CCDC152 expression acts as a negative feedback factor in GPCR via inhibition of Rho/ROCK signaling system.