The Study Of Mechanism Of Calcitriol Inhibit The Advanced Prostate Cancer

Objective: We use calcitriol to intervent DU145 and PC-3 prostate cancers by different concentration and time to assess the effect of proliferation and death performed by calcitriol, and explore the complicated mechanism.We want to employ the more rational project of combined drug therapy to optimize the anticancer efficacy. By this way, we expect to propose the better idea and threoy to support for advanced prostate cancer clinical therapy, and promote the new theory to translate to clinical application.Methods: We choose DU145 and PC-3 prostate cancer cell lines as cell models to evaluate the calcitriol anticancer effect. The two cell lines were cultured in vitro and amplified to suitable number.Then, we use different concentration of calcitriol 10 nM or 100 nM to intervent them respectively by indicated time point.We used CCK-8 detection method to assess the two cell proliferation rate of the control by different time point and different drug concentration, and assess the total cell death rate of the two cell lines by the trypan blue staining by the same condition. We applied the PI staining in the nucleus by FCM to evaluate the cell cycle of different group and applied the FCM to detect the Annexin V positive cell percentage hold by total cells to assess the early apoptosis percentage.We applied westernblot method to evaluate the key protein express level in the apotosis and antophagy signal pathway to judge the pathway activation state respectively. We used IHC and IF method to assess the VDR and LC3 location and expression. At last, we used chloroquine to block the autophagy signal pathway to evaluate whether the chloroquine can sensitivate the inhibitory effect of calcitriol to PC-3 cell.Results: Calcitriol can promote proliferation and inhibit apoptosis to DU145 cell by 72 hours and the interference to the autophagy is not significant. On the other hand,Calcitriol can obviously inhibit the proliferation and promote the total cell death subjected to PC-3 cell. In addition,we observe that the apparent sensitivity by chloroquine to calcitriol about the proliferation inhibition and cell death promotion which is attribute to the autophagic upstream activation and autophagic downstream inhibition simutaneously.Conclusion: Calcitriol can promote proliferation and inhibit apoptosis to DU145 cell. On the other hand, calcitriol can inhibit the PC-3 cell progression by activation of autophagy. Importantly, chloroquine has apparent sensitivate calcitriol as for PC-3 cell progression by the block of autophagic downstream process.