Lentivirus-Mediated Knockdown Of RTN4 In Prostate Cancer Inhibits Cell Proliferation,Cell Cycle And Induces Cell Senescence

Objective Prostate cancer is one of the most malignant tumor of the urinary and reproductive system,which is the most common types of cancer occuring to the men in the Europe and the USA,with the mortality rate only next to lung cancer. In the meanwhile, the incidence and the detection rate of prostate cancer are now rising in China every year.We have found that there are differences between the expression of RTN4(Reticulon-4) in prostate cancer and normal prostate epithelial tissues. However, the express and the effect of RTN4 in prostate cancer cell lines have not been studied yet.This research focus on the relationship between the RTN4 gene and the proliferation as well as the senescence of human prostate cancer cell in vitro.Methods(1)RT-PCR and Western blotting were used to detect the expression of the RTN4’s m RNA and protein in Prostate cancer cells,such as PC3,DU145 and LNCa P.(2) PTN4 specific si RNA vector was constructedthen the prostate cancer cells were infected,meanwhile untransfected group,control group and PTN4 si RNA transfected group were set up. Western blotting was performed to detect the interference effect.(3)MTT methodplate clone formation assay, FCM method, PI and SA-β-Ga1(senescence-associated beta-ga1actosidase) were used to detect the proliferation, cell cycle and the senescence of the prostate cancer cells.Results The protein and m RNA expression of PTN4 are both positive in PC3, DU145 and LNCa P. The expression of RTN4 at m RNA and protein levels was reduced significantly(P<0.01),suggesting that the RTN4-silencing cell line(LV-sh RTN4) was successfully established. The proliferation, cells’ number and its clone formation of LV-RTN4-RNA group was suppressed when compared with scrambled group and control group(P<0.01),which also caused the increase of S phase proportion and the senescense(P<0.01).Conclusion(1) The protein and m RNA expression of RTN4 are both positive in PC3, DU145 and LNCa P,suggesting that RTN4 may interact with the development of prostate cancer.(2)Silencing the RTN4 gene in PC3 suppresseed the proliferation of the cells and accelerated the senescence of the cells,suggesting that RTN4 has its role in the proatate cancer development and may be a potential tumor marker.