Proliferation Of Bladder Cancer Cell Lines Was Mediated By PLCΕ-PKCΑ Signal Pathway

PART ONEObjective: To investigate the effect of PKC isforms on bladder cancer proliferation.Methods: Bladder cancer BIU-87 cell lines were treated by PKC isforms special inhibitor (PKCαspecial inhibitor Safingol, PKCδspecial inhibitor Rottlerin) and PKC promoter PMA. Protein of down stream was detected by RT-PCR and Western. Cell viability was investigated using MTT assay, and cell cycle was analyzed by FCM.Results: Proliferation of baldder cancer cell BIU-87 cell lines were significant inhibited by PKCαspecial inhibitor Safingol and in dose-independent manner, and cells were arrested at G0/G1 phase, while PKCδspecial inhibitor Rottlerin didn't, and were obviously promote by PKC promoter PMA, and cells at in G2/M phase were increased after treated. Furthermore, our results showed that Expression of immediately gene c-fos and c-jun was related with PKCαactivity, but no related with PKCδactivity.Conclusion: PMA a PKC promoter could promote the bladder cancer cell proliferation, while Safingol a PKCαinhibitor could suppress the cell proliferation, showed the crucial role of PKCαin bladder cancer cell proliferation, and Rottlerin a PKCδinhibitor promoted cell proliferation, indicated that cell proliferation promoted by PMA maybe because PMA promote PKCαactivation. So we presumed that PKCαplay a crucial role in bladder cancer proliferation, cell viability was significant inhibited after PKCαactivity suppression, and this role maybe mediated by immediately gene c-fos and c-jun.PART TWOObjective: To investigated the role of PLCε-PKCαsignal pathway in bladder cancer cell proliferation.Methods: Bladder cancer BIU-87 cell lines were transfected with PLCεshRNA, expression of endogenous PLCεwas detected by RT-PCR, cell proliferation was analyzed by MTT assay, PKCαactivity was detected by Western, expression of c-jun and c-fos was detected by RT-PCR and Western; shRNA transfected cells were treated with PKC inhibitor and promoter, then proliferation was analyzed using MTT assay.Results: Expression of endogenous gene was suppressed by PLCεshRNA transfection; cell proliferation, PKCαactivity and expression of downstream protein c-fos and c-jun were significantly inhibited by PLCεshRNA transfection, cell cycle was arrested at G0/G1 phase; cell proliferation which was inhibited by PLCεshRNA transfection could be reversed by by PKC promoter PMA, but can not reversed if retreated with Safingol not Rottlerin.Conclusion: Cell proliferatin were promoted by PLCεand acitiviated PKCα, PKCαcould activated by PLCε, cell proliferation inhibited by PLCεgene silencing could reverse by PMA a PKC promoter, but it couldn't reverse while cell pretreated with PKCαinhibitor not PKCδinhibitor, this indicated that Safingol was a special inhibitor of PKCα. Moreover, expression of c-fos and c-jun was obviously inhibited by PLCεgene silencing, and the expression also positive correlation with PKCαactivation, these results indicated that bladder cancer cell proliferation promoted by PLCεmaybe mediated by PKCα. PLCε-PKCαsignal pathway play a crucial role in bladder cancer proliferation which mediated by immediately gene c-fos and c-jun, and related with cell cycle adjustion.