The Study For Mechanism Of Ribosome Biogenesis Mediated By NS For The Tumorigenesis And Tumor Progression Of Prostate Cancer

ObjectiveTo reveal themechanism of ribosome biogenesis mediated by NS for thetumorigenesis and tumor progression of prostate cancer MethodsRetrieve the expression level of ribosomal protein individually in PCa and BPH by using Oncomine database to find the abnormally ribosomal protein. Then we detected the abnormal ribosomal protein expression level in PCa and PBH by RT-qPCR;and then, we detected the protein level of the most differential expressed RPS and RPL by western blot, and analyzed the correlation of their expression level and the clinical variables of PCa.After down-regulation the NS gene in PC3 cells,we separated the ribosome 80 S, 60 S large subunit and 40 S small subunit of PC3 cell by using sucrose gradient centrifugation separation technology. The level of ribosome 80 S, 60 S large subunit and 40 S small subunit were detected by a UV detector for continuous absorbance measurements at 254 nm. Level of the ribosomal proteins of the ribosome subunit which is affected by NS were identified by RT-qPCR. Screening the most significant changed ribosomal protein and explore the role of the ribosomal protein in prostate cancer progression by vitro and vivo experiment.Using c- Myc high expression plasmid and c-Myc small molecule inhibitors 10058-F4 to processing PC3 cells and up-regulate the ROS level in PC3 cells by using H2O2 and down-regulate the ROS level by using catalase. then, we detected the NS expression level by RT-qPCR and western blot and the ROS level was detected by DCFH-DA probe. MTT and cell counting was used to detect the influence of c-Myc, ROS and NS on PC3 cells proliferation. ResultsBy searching the Oncomine database,we find there 15 RPs and 14 RPL were abnormally expressed in PCa, then,we detected their expression level in PCa and BPH of Chinese people.We find there 11 RPS and 11 RPL were abnormally expressed in PCa,and the most differential expressed RP is RPS2 and RPL36. We analyzed their relationship with clinical variables of PCa,we find that,the expression level of these two PR were closely related to the pathology and prognosis of PCa.The level of ribosome 60 S large subunit and 80 S polysome were reduced and the RPL19 mRNA level was changed most significantly after down-regulation the NS gene in PC3 cells. The protein level of RPL19 was consistent with NS. The cell proliferation and invasion ability in vitro and tumorigenesis ability in nude mice were discounted, the final tumor volume and weight were also decreased after down-regulation RPL19 gene.Conclusion Some RP were abnormally expressed in PCa, and the abnormally expressed RP is associated with a poor prognosis of PCa.Thec-Myc could elevate the ROS level and promote NS gene expression level in PC3 cells and ROS could up-regulate NS protein expression level. The cell proliferation was increased by up-regulate c-Myc and ROS in PC3 cells, while down-regulate NS expression level in PC3 cells could prohibiting the propagation of the PC3 cells. ConclusionThe c-Myc-ROS-NSmediated ribosomes biosynthesissignaling pathway is exist in the prostate cancer. Inhibition of this signaling pathway is a promising strategy to restrain the Pca progression.