Resaerch On Montelukast To The Apoptosis-Related Protein Of Castration-Resistant Prostate Cancer

Objective:The influence of Montelukast Sodium Hydrate on the proliferation reproduction, morphology and apoptosis of prostate cancer cell lines including PC3, DU145 and LNCap AND Homo sapiens prostate normal Epithelial cell line RWPE1 and the related mechanism were studied which might provide a new way of drug selection and preliminary experimental basis for castration-resistant prostate cancer therapy.Methods:The growth inhibition of MSH on PC3,DU145,LNCaP and RWPE1 Cell lines were test by Cell Counting Kit-8(CCK8) and the value of IC50 was calculated. After a series of concentration(0.3125,0.625,1.25,2.5,5,10,20ug/mL ) of Montelukast Sodium treatment, the changes of cell morphology was obserbed by light microscope and transmission electron microscopy. The apoptosis of prostate cancer cells was detected by flow cytometer. Additionally, the mRNA and proteins expression of WNT5A, β-catenin, E-cadherin genes were assessed by quantitative real-time fluorescence quantification and western blot.Results:(1) The results of CCK8 suggested that the values IC50 of Montelukast Sodium for DU145, PC3, LNCaP and RWPE-1 cell lines were about 2.7μg/ml, 3μg/ml,7μg/ml and 5μg/ml respectively.(2) In the light microscope, prostate cancer cells with no medicine was larger, core categories circular, seen nucleoli clearly, intercellular connection tightly.As for disposed group,prostate cancer cell was shrinkage and round, shedding. As for the transmission electron microscopy, the LNCaP and PC3 and DU145 cells appeared to be karyopyknosis,chromatin mass, side set clear and nuclear membrane is dissolved.The typical apoptosis body was also presented.(3) The results of flow cytometer identified that the prostate cancer cells was shown to be apoptotic in later period. Along with the increasing concentration and acting time of drug, the apoptosis of DU145, PC3 and LNCaP were elevated.(4) Comparing with blank control group, the expression of E-cadherin (P<0.05) gene in DU145, PC3 and LNCaP cell lines raised after qRT-PCR. WNT5A gene increased in DU145 (P<0.01) and decline in PC3 and LNCaP (P<0.05). As for β-catenin gene, they were just elevated in PC3 and LNCaP (P<0.05).(5) As for western blot, the protein expression of E-cadherin and WNT5A increased in DU145 cell line. And the expression of P-catenin descended and had statistical discrepancy in 3βg/ml group (P<0.01). In PC3 cell, the expression of E-cadherin (significant in 3βg/ml group P<0.05), WNT5A (significant in 1.5μg/ml group P<0.001) and β-catenin were decreasing.. In addition, in LNCaP, the decreasing expression was shown in E-cadherin, WNT5A (significant in 1.5μg/ml group P<0.05). On the opposite, just β-catenin was elevated with statistical discrepancy in the 3μg/ml group (P<0.01).Conclusion:MSH could restrain the development of prostate cancer cells and promote the occurrence of the late period apoptosis. Meanwhile, it could also disturb the Wnt/β-catenin pathway transition influencing expression of some related proteins. Considering the differences of effect of MSH on androgen-dependent and non-androgen-dependent cancer cells.