烟管头草水提物对前列腺癌PC3细胞增殖、迁移与侵袭的影响

Effects of Water Extract from Carpesium cernuum on Proliferation,Metastasis and Invasion of Prostate Cancer PC3 Cells

目的:研究烟管头草水提物(AECC)对前列腺癌PC3细胞增殖、迁移与侵袭的影响。方法:将细胞分为对照组和AECC不同质量浓度组(5、10、20、40、80μg/L),加入相应药物或培养基培养不同时间(24、48、72 h)后,检测细胞存活率。将细胞分为对照组和AECC低、中、高质量浓度组(20、40、80μg/L),同法培养24 h后,采用Hoechst 33258染色法和流式细胞仪检测各组细胞的凋亡情况;采用Transwell实验检测细胞迁移数和侵袭数;采用实时荧光定量聚合酶链式反应和Western blot法检测AECC低、中质量浓度组细胞中β-连环蛋白(β-catenin)信号通路相关迁移与凋亡蛋白[β-catenin、基质金属蛋白酶7(MMP-7)、髓细胞瘤病毒致癌基因(c-Myc)、胱天蛋白酶3(caspase-3)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)]的mRNA和蛋白表达水平。结果:随着给药浓度的增加和作用时间的延长,AECC不同质量浓度组细胞存活率均显著低于对照组(P<0.05或P<0.01),且呈不断降低趋势。与对照组比较,AECC各质量浓度组细胞的早期凋亡率(中质量浓度组除外),细胞迁移数和侵袭数,以及AECC低、中质量浓度组细胞中MMP-7、c-Myc(低质量浓度组除外)、Bcl-2(低质量浓度组mRNA除外)的mRNA和蛋白表达水平均显著降低(P<0.05或P<0.01);AECC各质量浓度组细胞的晚期凋亡率,AECC低、中质量浓度组细胞中β-catenin、caspases-3(低质量浓度组除外)、Bax(低质量浓度组mRNA除外)的mRNA和蛋白表达水平均显著升高(P<0.05或P<0.01)。结论:AECC可抑制PC3细胞的增殖、迁移与侵袭,其作用机制可能与调控β-catenin信号通路相关的迁移与凋亡因子的表达有关。

OBJECTIVE:To study the effects of the water extract from Carpesium cernuum(AECC)on the proliferation,metastasis and invasion of prostate cancer PC3 cells.METHODS:Cells were divided into control group and different concentration groups of AECC(5,10,20,40,80μg/L),and then treated with relevant medicine or medium for different time(24,48,72 h).The survival rates of cells were detected.Cells were divided into control group,and AECC low,medium and high concentration groups(20,40,80μg/L).After cultured for 24 h,Hoechst 33258 staining and flow cytometry were used to detect the apoptosis of cells.The number of cell metastasis and invasion were detected by Transwell assay.RT-qPCR and Western blot assay were applied to detect the mRNA and protein expression ofβ-catenin signaling pathway related migration and apoptosis proteins(β-catenin,MMP-7,c-Myc,caspase-3,Bcl-2 and Bax)in AECC low and medium concentration groups.RESULTS:With the increase of the concentration and culture time,the survival rates of cells in AECC different concentration groups were significantly lower than control group(P<0.05 or P<0.01),and showed a decreasing trend.Compared with control group,the early apoptosis rate(except the medium concentration group)and the number of cell metastasis and invasion in AECC groups,the mRNA and protein expression of MMP-7,c-Myc(except for the low concentration group)and Bcl-2(except for mRNA of the low concentration group)in AECC low and medium concentration groups were decreased significantly(P<0.05 or P<0.01).Late apoptosis rate of AECC groups,the mRNA and protein expression ofβ-catenin,caspases-3(except for the low concentration group),Bax(except for mRNA of the low concentration group)in AECC low and medium concentration groups were increased significantly(P<0.05 or P<0.01).CONCLUSIONS:AECC could inhibit the proliferation,metastasis and invasion of PC3 cells;the mechanism of which may be associated with regulating the expression ofβ-catenin signaling pathway related migration and apoptotic factors.