Study The Effect Of Osthole On Human Prostate Cancer PC-3 Cell In Vitro

Objective: Prostate cancer has developed into one of the most common cancers in men,it has been a serious threat to the health of old male. Because of lacking of typical symptoms in early stage,when the disease was diagnosed,it is already in T3 or T4 even later,missing the best opportunity of operation.At present,androgen blocking method is the main method in the treatment of limitations of advanced prostate cancer and recurrence,metastasis of prostate cancer.But With the extension of time that majority of patients treated with with the hormone blocking method,the disease progressing to androgen independent prostate cancer,and we lack of effective treatment measures to it.Osthole is a coumarin, which is a kind of extraction from Fructus Cnidii of biologically active substances,it has a significant effect in the treatment of various malignant tumors,but at present,our study is limited in the treatment and prevention of prostate cancer.The purpose of this experiment is to study the effects of Fructus Cnidii on the proliferation,cell cycle and apoptosis of Androgen independent prostate cancer PC-3 cells to investigate the mechanism and the potential of new drugs.Methods: Using cell culture techniques of cultured human prostate cancer cell line PC-3 in vitro,using different concentrations of osthole drug intervention,and the control group was established. after the intervention of different concentrations of osthole on human prostate cancer cell line PC-3 by MTT, observing the proliferation of each PC-3 changes with time,After intervention the various concentrations of osthole on experimental group PC-3 cells by Flow cytometry,observing the distribution of cell cycle and apoptosis,After detection of osthole intervention of PC-3 prostate cancer cell by western blot,observing the expression of subunits P110 and p85 of PI3 K and the expression level of P-Akt.The experimental data were analyzed by SPSS13.0 statistical software, P < 0.05 means statistical difference,P < 0.01 is significant difference.Results:1 MTT analysis results showed that,osthole could inhibit the proliferation on human prostate cancer PC-3 cell,with the time prolonging of drug intervention and the increase of drug concentration,the inhibition rate is growing up,(P<0.01), the results is in a concentration and time dependent manners.2 Flow cytometry analysis results suggested that, after each concentration group of osthole intervention,PC-3 prostate cancer cell line G0/G1, S phase cells decreased significantly,G2/M phase cells increased,means that there was a statistical significance(P<0.05),proving that Osthol could produce G2/M phase arrest. The cells apoptosis of experimental group was inspected by flow cytometryrate,that is2.87%, 10.82%, 22.05% and 33.33%.In addition to the 6.25 M data sets with no statistical difference compared with the control group(P>0.05), the other groups had statistically significant(P<0.05), the experimental group is further multiple comparisons,the difference was statistically significant(P<0.05), namely the osthol coud induce apoptosis of PC-3 cell,With increasing of concentration of osthole, the effect is more obvious.3 The resulets of Western blotting showed that, osthole intervention of PC-3 prostate cancer cell, the expression level of Phosphorylation of Akt in the PI3K/Akt signaling pathway and PI3 K two subunits P110, p85 had been decreased.compared with the control group, the difference was statistically significant(P<0.05). Each experimental group in relation to each other, The expression level of P110, p85 and P-Akt decreased gradually with the increase of concentration with significant difference(P<0.05). Osthole can downregulate the the expression level of P-Akt and P110 and p85,The expression level of P-Akt and P110 and p85 of osthole concentration tented to be inversely related.Conclusion:1 Osthole inhibits the proliferation on human prostate cancer cell line PC-3 in vitro,showed a dose,time dependence.Human prostate cancer cell line PC-3 were arrested in G2/ M phase.2 Osthole can induce the apoptosis of human prostate carcinoma PC-3 cells,and show time,dose dependent relationship,The mechanism may be related with decreasing the expression level of P-Akt in the PI3K/Akt signaling pathway and P110, p85,the two subunits of PI3 K.So we can infering that blocking the PI3K/ARK signaling pathway in PC-3 cells is one of the mechanism of the action of osthole.