白杨素调控Wnt/β-catenin通路对前列腺癌PC-3细胞株侵袭和迁移的影响

Effect of Chrysin Regulating Wnt/β-catenin Pathway on Invasion and Migration of Prostate Cancer Cell Line PC-3

目的:探讨白杨素调控Wnt/β-连环蛋白(β-catenin)通路对前列腺癌PC-3细胞株侵袭和迁移的影响。方法:设PC-3细胞组、5-氟尿嘧啶组、白杨素低剂量组、白杨素高剂量组。5-氟尿嘧啶组加入5-氟尿嘧啶使最终浓度为80.0μg/mL,白杨素低、高剂量组加入白杨素使最终浓度分别为60.0μg/mL、120.0μg/mL,培养72 h。四甲基偶氮唑盐微量酶反应比色法(MTT)及结晶紫染色测定细胞存活率及克隆形成数目,Transwell法测定细胞侵袭水平,annexinV-FITC/PI试剂盒测定细胞凋亡水平,逆转录聚合酶链式反应(RT-PCR)法及蛋白质印迹法(Western blot)测定细胞β-catenin基因和蛋白水平。结果:与PC-3细胞组比较,其余3组细胞存活率、克隆形成数目、穿膜数、β-catenin mRNA和蛋白表达水平均降低(P<0.05),凋亡率升高(P<0.05)。与白杨素低剂量组比较,白杨素高剂量组细胞、存活率、克隆形成数目、穿膜数、β-catenin mRNA和蛋白表达水平降低(P<0.05),凋亡率升高(P<0.05)。与5-氟尿嘧啶组比较,白杨素低剂量组细胞、存活率、细胞克隆形成数目、穿膜数、β-catenin mRNA和蛋白表达水平升高(P<0.05),凋亡率降低(P<0.05);白杨素高剂量组细胞、存活率、细胞克隆形成数目、穿膜数、β-catenin mRNA和蛋白表达水平降低(P<0.05),凋亡率升高(P<0.05)。结论:白杨素能抑制前列腺癌PC-3细胞株增殖、侵袭,诱导前列腺癌PC-3细胞株凋亡,其机制与白杨素可通过抑制β-catenin mRNA及蛋白的表达进而抑制Wnt/β-catenin通路有关。

Objective: To discuss the effect of chrysin regulating Wnt/β-catenin(β-catenin) pathway on invasion and migration of prostate cancer cell line PC-3. Methods: There were four groups set, namely the cell PC-3 group, the 5-fluorouracil group, the low-dose chrysin group and the high-dose chrysin group. In the 5-fluorouracil group, the final concentration was up to 80.0 μg/mL after adding 5-fluorouracil;in the low-dose chrysin group and the high-dose chrysin group,the final concentration was 60.0 μg/mL and 120.0 μg/mL respectively after adding chrysin;these four groups were cultivated for 72 hours. The survival rate and the number of colony formation of cells were detected via methyl thiazolyl tetracolium(MTT) and crystal violet staining. The level of cell invasion was detected via Transwell method. The level of cell apoptosis was detected via annexinV-FITC/PI. The levels of gene and protein of β-catenin were detected via reverse transcriptase polymerase chain reaction(RT-PCR) and Western blot. Results:Compared with those in the cell PC-3 group,the survival rate, the number of colony formation, the number of cell invasion, and the expression levels of β-catenin mRNA and protein in the other three groups were decreased(P<0.05),and the rate of apoptosis were increased(P<0.05).Compared with those in the low-dose chrysin group, in the high-dose chrysin group, the survival rate, the number of colony formation,the number of cell invasion,and the expression levels of β-catenin mRNA and protein were decreased(P<0.05),and the rate of apoptosis was increased(P<0.05). Compared with those in the 5-fluorouracil group,in the lowdose chrysin group, the survival rate, the number of colony formation, the number of cell invasion, and the expression levels of β-catenin mRNA and protein were increased(P<0.05),and the rate of apoptosis was decreased(P<0.05);in the high-dose chrysin group, the survival rate, the number of colony formation, the number of cell invasion, and the expression levels of β-catenin mRNA and protein were decreased(P<0.05), and the rate of apoptosis was increased(P<0.05). Conclusion:Chrysin can inhibit the proliferation and invasion of prostate cancer cell line PC-3 and induce its apoptosis,whose mechanism is related to the inhibition of chrysin on the mRNA and protein expression of β-catenin and the further inhibition on Wnt/β-catenin pathway.