NOP16 Lower Expression Enhance Chemotherapy Sensitivity In Prostate Cancer

Objective :To confirm relations of expression degree of NOP16 in prostate cancer with transform about biological characteristics of prostate cancer cell,and detect relations of degree of NOP16 and docetaxel chemosensitivity to prostate cancer cell and prostate cancer in nude mouse.And detect biological mechanism of low expression degree of NOP16.Methods : We design the RT-qPCR to measure m RNA expression count in prostate cancer sample and BPH sample.We design the RT-q PCR and Western Blot to measure m RNA expression count and protein count of NOP16 in the cell line of RWPE-1,LNCa P and PC3 m RNA expression count in prostate cancer sample and BPH sample.We design test to get low expression of NOP16 through NOP16 sh RNA and PC3 cell,we call it PC3-NOP16-KD.Then we test m RNA expression count and protein count of NOP16 in transduced PC3 cells with RT-q PCR and Western Blot.CCK-8 assay and cell count test was be used to test proliferation of NOP16 low expressed cells.Transwell assay and annexin V assay were used to estimate invasion ability and apoptosis ability of NOP16 expressed decreased transduced PC3 cells.The total amount of cell synthetic protein was be counted response unusual NOP16 expression on protein synthesis.CCK-8 assay was used to detect PC3 cell activation with different docetaxel concentrations sustain 24 h,48h;Docetaxel sub-lethal amount to PC3 cell deal with PC3-Vector cell?PC3-NOP16-KD cell then test cell activation.Mon-clone assay with PC3-Vector cell and PC3-NOP16-KD cell dealed with docetaxel were used to test cell proliferation ability;Cultivate PC3-Vector cell and PC3-NOP16-KD cell and dealed with docetaxel,then apoptosis assay were used to test cell apoptosis;Cultivate PC3-Vector cell and PC3-NOP16-KD cell and dealed with docetaxel,then cell cycle assay were used to test cell cycle;Cultivate PC3-Vector cell and PC3-NOP16-KD cell and dealed with docetaxel,then transwell assay were used to measure cell invasive ability.Western Blot assay was used to test Caspase3 in PC3 cell,PC3-Vector cell and PC3-NOP16-KD cell,these samples were dealed with 0nmol/L,5nmol/L,10nmol/L concentrations docetaxel sustain 24 h.We analyzed these data with SPSS 18.0 statistical software.Results : NOP16 is higher expressing in prostate cancer sample than that in benign prostatic hyperplasia sample;NOP16 are higher expressing in LNCa P and PC3 cell lines than that in the RWPE-1cell line;And similar expression of NOP16 were detected from LNCa P and PC3 cell lines.Established lower expression of NOP16 into PC3 cells,by specific sh RNA.Cell proliferation decreasing following lower expression of endogenous NOP16.Cell invasion decreasing following lower expression of endogenous NOP16.Increase cell apoptosis following lower expression of endogenous NOP16.Lower cellular protein synthes following lower expression of endogenous NOP16.Cell activity decreasing when PC3 cell were dealed with different concentration docetaxel.IC50 about 30nmol/L(24h),20nmol/L(48h);PC3-Vector cell and PC3-NOP16-KD cell were dealed with sub-lethal amount docetaxel.Result display,following docetaxel concentration elevating cell activity obvious decreasing.NOP16-KD cell were more sensitivity to Docetaxel,NOP16 elevate docetaxel chemosensitivity to prostate cancer cell.NOP16-KD group clones were obvious fewer than control group,NOP16 resist to cell damage effect of docetaxel.NOP16-KD group cell apoptosis were higher than control group cell,and prostate cancer cell apoptosis were higher than dealed with docetaxel group,lower NOP16 expression elevate Docetaxel chemosensitivity.Cell cycle assay confirmed that cell proliferation ability in NOP16-KD group is lower than control group;And dealed with docetaxel proliferation ability in NOP16-KD group is lower than control group.NOP16 resist inhibiting effect to cell of docetaxel;Transwell assay showed lower invasiveness ability in NOP16-KD group cell than that in control group cell.NOP16 resist inhibiting effect to cell of docetaxel through affect invasiveness ability;Compared to control group cell,Caspase3 were higher after dealed with docetaxel,These showed that NOP16 induce prostate cancer cell apoptosis cooperate with Docetaxel.In asaay of tumor formation in nude mice,bigger tumor in NOP16-KD group cell than in control group cell;Treating with docetaxel tumor were further shrink in NOP16-KD grou.NOP16 obvious enhance docetaxel chemosensitivity to prostate cancer in mice.Compared NOP16-KD-DOX group to PC3-DOX group,many differential genes were found through gene chip experiment.Conclsion: Abnormal expression of NOP16 in prostate cancer cell change biological behaviour in vitro and vivo.NOP16 obvious enhance docetaxel chemosensitivity to prostate cancer in vitro and vivo.We believe NOP16 will be potential therapeutic target of prostate cancer.