三七总皂苷对前列腺癌PC-3细胞增殖及迁移的抑制作用

摘要

目的:探讨三七总皂苷对人前列腺癌PC-3细胞增殖及迁移的抑制作用,初步研究该药物的作用机制,并为前列腺癌的药物治疗及扩展tPNS的临床应用提供实验依据.方法:采用MTT实验、细胞计数、伤口愈合实验等方法检测tPNS对PC-3细胞增殖及迁移的影响;采用Western blot及明胶酶图分析等方法对增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、血管细胞间黏附分子1(vascular cell adhesion molecule 1,VCAM-1)及迁移相关蛋白基质金属蛋白酶2(matrix metalloproteinases 2,MMP-2)等进行检测.通过检测p38 MAPK及ERK途径的磷酸化变化,探讨tPNS对PC-3细胞作用的可能信号途径.结果:tPNS可抑制PC-3细胞增殖,随着tPNS浓度(200、400、800 mg/L)的增加,对PC-3细胞的抑制率分别增加至13.0%、29.5%和35.9%(P＜0.05).tPNS下调PC-3细胞增殖标志物PCNA的表达水平,该效应呈量效及时效关系.tPNS(200、400、800 mg/L)可显著抑制PC-3细胞迁移.tPNS还可显著下调迁移相关蛋白MMP-2及黏附分子VCAM-1的表达水平.tPNS可显著增加p38 MAPK的磷酸化水平,但对ERK磷酸化影响不明显.结论:tPNS抑制人前列腺癌PC-3细胞增殖及迁移活性,这些生物学作用可能与该药抑制PCNA、VCAM-1、MMP-2的表达及激活p38 MAPK信号通路有关.%Objective: To investigate the effects of total Panax notoginseng saponins ( tPNS ) on the proliferation and migration of PC-3 cells, and to provide experimental evidence for the use of tPNS in treating prostatic cancer.Methods: MTT, cell counting, and wound-healing assays were performed to analyze the effects of tPNS on the proliferation and migration of PC-3 cells.The expression of proliferating cell nuclear antigen ( PCNA ), vascular cell adhesion molecule 1 ( VCAM-1 ), and matrix metalloproteinases 2 ( MMP-2 ) was determined using Western blot analysis or zymography.Activation of p38 MAPK and ERK was also detected by Western blot analysis after tPNS treatment.Results: tPNS inhibited the proliferation of PC-3 cells.The inhibition ratios were 13.0％, 29.5％, and 35.9％ after the PC-3 cells were treated with tPNS ( 200, 400, and 800 mg/L, respectively ) ( P ＜ 0.05 ).PCNA expression was also inhibited in dose- and time-dependent manners.These results indicated that tPNS could significantly inhibit PC-3 cell proliferation via downregulation of PCNA.The migration of PC-3 cells was inhibited after treatment with tPNS.The Western blot and zymography results suggested that tPNS significantly suppressed the expression of the migration-related protein MMP-2.VCAM-1 was also downregulated after tPNS treatment in PC-3 cells.The aforementioned results indicated that tPNS inhibited PC-3 cell migration by suppressing MMP-2 and VCAM-1 expression.The phosphorylation level of the p38 MAPK pathway was dramatically increased at 0.5, 1, 3, and 6 h after tPNS treatment ( 400 mg/L ).However, phosphorylation of the ERK pathway was unaffected.Conclusion: tPNS can inhibit the proliferation and migration of PC-3 cells.The underlying mechanism may be related to the expression of PCNA, VCAM-1, and MMP-2 and activation of the p38 MAPK pathway.