三七总皂苷调控巨噬细胞对乳腺癌细胞MDA-MB-231侵袭迁移影响

目的:探讨三七总皂苷(Panax notoginseng saponins,PNS)通过调控巨噬细胞对乳腺癌细胞MDA-MB-231侵袭迁移的影响。方法:培养MDA-MB-231和Raw264.7细胞,给予不同浓度PNS,采用MTT与流式细胞术(Flow Cytometry,FCM)分别检测不同浓度PNS对MDA-MB-231与巨噬细胞Raw264.7增殖与凋亡的影响;划痕实验检测不同浓度PNS处理的Raw264.7细胞的条件培养基对乳腺癌细胞MDA-MB-231迁移的影响;Transwell实验检测不同浓度PNS与巨噬细胞共同作用对乳腺癌细胞MDA-MB-231侵袭的影响;ELISA与流式细胞术检测不同浓度PNS对巨噬细胞相关细胞因子分泌及蛋白表达的影响。结果:PNS可以剂量依赖性地抑制MDA-MB-231与Raw264.7细胞的增殖,高浓度PNS溶液可以显著增加细胞凋亡;低浓度(20,50μg·mL^-1)PNS巨噬细胞条件培养基可以抑制MDA-MB-231的迁移;低浓度(20,50μg·mL^-1)PNS与巨噬细胞共培养可以明显抑制MDA-MB-231的侵袭;低浓度(20,50μg·mL^-1)PNS促进巨噬细胞IL-6与TNF-α的分泌,并上调巨噬细胞表面CD86的表达水平。结论:PNS可通过调控巨噬细胞抑制乳腺癌细胞MDA-MB-231侵袭、迁移,并可通过调控巨噬细胞向M1型巨噬细胞极化,进而发挥抑制肿瘤的作用。

Panax notoginseng saponins regulates macrophage invasion of MDA-MB-231 in breast cancer cells migration effects

OBJECTIVE To investigate the effect of Panax notoginseng saponinson the invasive migration of breast cancer cells MDA-MB-231 by regulating macrophages.METHODS MDA-MB-231and Raw264.7cells were cultured and given different concentrations of PNS,and the effects of different concentr ations of Panax notoginseng saponins(PNS)on the proliferation and apoptosis of MDA-MB-23 and macrophages Raw264.7 were detected by MTT and flow cytometry(FCM),respectively;scratch assay was performed to detect the effects of different concentrations of PNS treated Raw264.7cells treated with different concentrations of PNS on the migration of MDA-MB-231 breast cancer cells;Transwell assay to detect the effect of different concentrations of PNS and macrophages on the invasion of MDA-MB-231breast cancer cells;ELISA and flow cytometry to detect the effect of different concentrations of PNS on macrophage-related cytokine.The effects of PNS on macrophage-related cytokine secretion and protein expression were examined by ELISA and flow cytometry.RESULTS PNS dose-dependently inhibited the proliferation of MDA-MB-231 and Raw264.7 cells,and high concentrations of PNS solution significantly increased apoptosis;low concentrations(20,50 μg·mL^-1)of PNS macrophage conditioned medium inhibited the migration of MDA-MB-231.Low concentrations(20,50 μg·mL^-1)of PNS co-cultured with macrophages significantly inhibited the invasion of MDA-MB-231;low concentrations(20,50 μg·mL^-1)of PNS promoted the secretion of IL-6 and TNF-α in macrophages and up-regulated the expression level of CD86 on the surface of macrophages.CONCLUSION PNS can inhibit the invasion and migration of breast cancer cells MDA-MB-231 by regulating macrophages,and can exert tumor suppressive effects by regulating the polarization of macrophages to M1-type macrophages.