胡黄连苷Ⅱ对非小细胞肺癌恶性进展的影响

Effect of picrosideⅡon the malignant progression of non-small cell lung cancer

目的探讨胡黄连苷Ⅱ对非小细胞肺癌(NSCLC)恶性进展的影响及机制。方法将A549细胞分组为对照组,胡黄连苷Ⅱ低、中、高浓度组,K6PC-5[鞘氨醇激酶1(SPHK1)激活剂]组,胡黄连苷Ⅱ高剂量+K6PC-5组,检测细胞增殖、迁移、侵袭情况,以及细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶2(MMP-2)、MMP-9、SPHK1、1-磷酸鞘氨醇受体3(S1PR3)及胞外信号调节激酶1/2(ERK1/2)蛋白的表达情况。以BALB/c裸鼠为对象,通过皮下接种A549细胞悬液建立NSCLC裸鼠移植瘤模型,并将其分为裸鼠-对照组,裸鼠-胡黄连苷Ⅱ低、中、高剂量组,裸鼠-K6PC-5组,裸鼠-胡黄连苷Ⅱ高剂量+K6PC-5组(每组5只),考察胡黄连苷Ⅱ对其瘤体质量及体积的影响。结果与对照组比较,胡黄连苷Ⅱ低、中、高浓度组的细胞OD450值、EdU阳性细胞率、划痕愈合率、细胞侵袭数及PCNA、MMP-2、MMP-9、SPHK1、S1PR3、ERK1/2蛋白的相对表达量均显著降低;与裸鼠-对照组比较,裸鼠-胡黄连苷Ⅱ低、中、高剂量组裸鼠体内的瘤体质量及体积均显著降低或缩小,上述指标均呈浓度/剂量依赖性变化(P<0.05);K6PC-5组细胞和裸鼠-K6PC-5组裸鼠对应指标的变化趋势则相反(P<0.05)。与胡黄连苷Ⅱ高浓度组或裸鼠-胡黄连苷Ⅱ高剂量组比较,胡黄连苷Ⅱ高浓度+K6PC-5组细胞和裸鼠-胡黄连苷Ⅱ高剂量+K6PC-5组裸鼠的上述定量指标均显著升高或增大(P<0.05)。结论胡黄连苷Ⅱ可能通过抑制SPHK1/1-磷酸鞘氨醇/S1PR3信号通路来抑制NSCLC的恶性进展。

OBJECTIVE To investigate the effect and mechanism of picrosideⅡon the malignant progression of non-small cell lung cancer(NSCLC).METHODS A549 cells were divided into the control group,picrosideⅡlow-,medium-and highconcentration groups,K6PC-5[sphingosine kinase 1(SPHK1)activator]group,and picrosideⅡhigh-dose+K6PC-5 group.Cell proliferation,migration and invasion were detected.Besides,the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),MMP-9,SPHK1,sphingosine-1-phosphate receptor 3(S1PR3)and extracellular signal-regulated kinase 1/2(ERK1/2)protein in the cells were also observed.BALB/c nude mice were subcutaneously inoculated with A549 cell suspension to establish NSCLC xenograft models.Then they were assigned to the nude mouse-control group,nude mouse-picrosideⅡlow-,medium-and high-dose groups,nude mouse-K6PC-5 group,and nude mouse-picrosideⅡhigh-dose+K6PC-5 group(with 5 mice in each group)to investigate the effect of picrosideⅡon their tumor mass and volume.RESULTS Compared with the control group,the OD450 values,EdU-positive cell rates,scratch healing rates,cell invasion number,and the relative expression levels of PCNA,MMP-2,MMP-9,SPHK1,S1PR3 and ERK1/2 protein in the low-,medium-and high-concentration groups of picrosideⅡwere significantly decreased.Compared with the nude mouse-control group,the tumor mass and volume in the nude mouse-low-,medium-and high-dose groups of picrosideⅡwere significantly decreased or shrunk.The changes of above indicators were concentration/dose-dependent(P<0.05).The changing trend of the corresponding indicators in the K6PC-5 group and the nude mouse-K6PC-5 group was opposite(P<0.05).Compared with the picrosideⅡhigh-concentration group or the nude mice-picrosideⅡhigh-dose group,the above quantitative indicators in the picrosideⅡhigh-concentration+K6PC-5 group cells and the nude mouse-picrosideⅡhigh-dose+K6PC-5 group nude mice were significantly increased or enlarged(P<0.05).CONCLUSIONS PicrosideⅡmay inhibit the malignant progression of NSCLC by inhibiting SPHK1/sphingosine-1-phosphate/S1PR3 signaling pathway.