三七总皂苷对前列腺癌PC-3细胞增殖及迁移的抑制作用

Inhibition of the Proliferation and Migration of Human Prostate Cancer PC-3 Cells by Total Panax Notoginseng Saponins

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、800mg/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、800mg/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 ceils. 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 downregula- tion 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 activa- tion of the p38 MAPK pathway.