欧前胡素对人前列腺癌PC3细胞增殖凋亡及RhoA/ROCK1信号通路的影响

Effects of Imperatorin on Proliferation,Apoptosis and RhoA/ROCK1 Signaling Pathway in Human Prostate Cancer PC3 Cells

目的:探讨欧前胡素对人前列腺癌PC3细胞增殖、凋亡及Ras同源基因家族成员A(RhoA)/Rho关联含卷曲螺旋结合蛋白激酶1(ROCK1)信号通路的影响。方法:设PC3细胞组、5-氟尿嘧啶组(100μg/mL)、欧前胡素低、高剂量组(60μg/mL、120μg/mL),以上各组每孔设6个平行样,培养72h。噻唑蓝(MTT)法测定细胞增殖,Transwell腔室系统测定细胞侵袭迁移水平,流式细胞仪测定细胞凋亡水平,实时荧光定量PCR(qRT-PCR)法及蛋白免疫印迹法测定细胞RhoA、ROCK1 mRNA和蛋白水平。结果:与PC3细胞组比较,5-氟尿嘧啶组、欧前胡素低、高剂量组OD值、存活率、穿膜数、RhoA、ROCK1 mRNA和蛋白表达水平降低,凋亡率升高(P<0.05);与5-氟尿嘧啶组比较,欧前胡素低、高剂量组OD值、存活率、穿膜数、RhoA、ROCK1 mRNA和蛋白表达水平升高,凋亡率降低(P<0.05);与欧前胡素低剂量组比较,欧前胡素高剂量组OD值、存活率、穿膜数、RhoA、ROCK1 mRNA和蛋白表达水平降低,凋亡率升高(P<0.05)。结论:欧前胡素能明显抑制人前列腺癌PC3细胞增殖、侵袭迁移,促进其凋亡,其机制可能与欧前胡素抑制人前列腺癌PC3细胞RhoA、ROCK1 mRNA和蛋白表达进而抑制RhoA/ROCK1信号通路的激活有关。

Objective:To investigate the effect of imperatorin on proliferation,apoptosis and Ras homolog gene family member A(RhoA)/Rho-associated coiled coil-containing protein kinase 1(ROCK1)signaling pathway in human prostate cancer PC3 cells.Methods:PC3 cell group,5-fluorouracil group(100μg/mL),low-dose and high-dose imperatorin group(60μg/mL,120μg/mL)were set up.Six parallel samples were set in each well of the above groups for 72 h.Cell proliferation was measured by MTT assay,and cell invasion and migration was measured by Transwell chamber system,apoptosis measured by flow cytometry,RhoA and ROCK1 mRNA and protein levels measured by qRT-PCR and Western blot.Results:Compared with PC3 cell group,the OD value,survival rate,penetration number,RhoA,ROCK1 mRNA and protein expression levels in 5-fluorouracil group,low-dose and high-dose imperatorin group were decreased,the apoptosis rate were increased(P<0.05).Compared with 5-fluorouracil group,the OD value,survival rate,penetration number,RhoA,ROCK1 mRNA and protein expression levels in low-dose and high-dose imperatorin group were increased,the apoptosis rate were decreased(P<0.05).Compared with low-dose imperatorin group,the OD value,survival rate,penetration number,RhoA,ROCK1 mRNA and protein expression levels in high-dose imperatorin group were decreased,the apoptosis rate were increased(P<0.05).Conclusion:Imperatorin can significantly inhibit the proliferation,invasion and migration,promote its apoptosis of human prostate cancer PC3 cells.The mechanism may be related to the inhibition of RhoA,Rock 1 mRNA and protein expression by imperatorin and then inhibiting the activation of RhoA/ROCK1 signaling pathway.