摘要
目的探讨柴胡皂苷D通过JAK2/STAT3通路对乳腺癌MCF-7细胞增殖和凋亡的影响。方法将乳腺癌MCF-7细胞分为对照组(0μmol/L),低(5μmol/L)、中(10μmol/L)和高(20μmol/L)剂量柴胡皂苷D组,采用四甲基偶氮唑盐(tetramethylazolium salt,MTT)比色法测定各组MCF-7细胞活力;用流式细胞术检测各组细胞凋亡情况;用免疫组化检测柴胡皂苷D对MCF-7细胞JAK2和STAT3表达情况;同时用免疫蛋白印迹法检测各组细胞JAK2、STAT3和Ki-67蛋白的表达。结果对照组、低剂量组、中剂量组和高剂量组MCF-7细胞活力差异有统计学意义(F=5.099、38.908、159.680、234.885、344.866、386.914,P<0.05)。与对照组相比较,在12、24、36、48、60、72 h后,低剂量组、中剂量组和高剂量组MCF-7细胞活力均有显著降低(P<0.05),且低剂量组细胞活力在24、36 h均高于中剂量组和高剂量组(P<0.05),中剂量组细胞活力在24、36 h均高于高剂量组(P<0.05)。对照组、低剂量组、中剂量组和高剂量组MCF-7细胞凋亡率比较差异具有统计学意义(χ^(2)=21.609,P<0.05);与对照组比较,低剂量组、中剂量组和高剂量组的细胞凋亡率均升高(P<0.05),低剂量组细胞凋亡率低于中剂量组和高剂量组(P<0.05),中剂量组细胞凋亡率低于高剂量组(P<0.05)。免疫组化结果显示,对照组、低剂量组、中剂量组和高剂量组MCF-7细胞JAK2、STAT3蛋白相对表达量比较差异具有统计学意义(F=44.343、46.389,P<0.05);与对照组比较,低剂量组、中剂量组和高剂量组MCF-7细胞JAK2、STAT3表达均降低(P<0.05),且低剂量组细胞JAK2、STAT3表达均高于中剂量组和高剂量组(P<0.05),中剂量组细胞JAK2、STAT3表达均高于高剂量组(P<0.05)。Western blot检测结果显示,对照组、低剂量组、中剂量组和高剂量组MCF-7细胞JAK2、STAT3和Ki-67蛋白相对表达量比较差异具有统计学意义(F=25.241、18.102、14.997,P<0.05);与对照组比较,低剂量组、中剂量组和高剂量组的细胞JAK2、STAT3和Ki-67相对表达量均降低(P<0.05),低剂量组细胞JAK2、STAT3相对表达量均高于中剂量组和高剂量组(P<0.05),中剂量组细胞JAK2、STAT3相对表达量均高于高剂量组(P<0.05)。结论柴胡皂苷D能够抑制乳腺癌MCF-7细胞增殖,促进MCF-7细胞凋亡,其作用机制可能与JAK2/STAT3通路有关。
Objective To investigate the effect of saikosaponin D on the proliferation and apoptosis of breast cancer MCF-7 cells by JAK2/STAT3 pathway.Methods The breast cancer MCF-7 cells were divided into control group,low-dose group,middle-dose,and high-dose groups(treated with 0,5,10 and 20μmol/L saikosaponin D for 12,24,36,48,60 or 72 h).Tetramethylazolium salt(MTT)colorimetric assay was used to detect the cell viability in MCF-7 cells of each group.Flow cytometry was adopted to detect cell apoptosis in each group.Immunohistochemical assay was employed to detect the expression of JAK2 and STAT3 in MCF-7 cells.At the same time,Western blotting was used to detect the protein levels of JAK2,STAT3 and Ki-67 in each group of cells.Results There was statistical differences in MCF-7 cell viability among the control,and low-,middle-and high-dose groups(F=5.099,38.908,159.680,234.885,344.866,386.914,P<0.05).Compared with the control group,the cell viability of MCF-7 cells in the low-dose,middle-dose and high-dose groups were significantly reduced after treatment for 12,24,36,48,60 and 72 h(P<0.05),and the cell viability of the cells in the low-dose group for 24 and 36 h was higher than that of the middle-dose group and the high-dose group(P<0.05),and the cell viability of the middle-dose group was higher than that of the high-dose group at 24 and 36 h(P<0.05).The apoptotic rate of MCF-7 cells differed significantly among the control group,and low-,middle-and high-dose groups(Chi-square=21.609,P<0.05).The rate was higher in the 3 treatment groups than the control group(P<0.05),that of the low-dose group was lower than those of the middle-dose group and the high-dose group(P<0.05),so was that of the middle-dose group lower than that in the high-dose group(P<0.05).The relative expression levels of JAK2 and STAT3 proteins in MCF-7 cells were significantly different among the 4 groups(F=44.343,46.389,P<0.05),they were obviously lower in the 3 treatment groups than the control group(P<0.05),and those in the low-dose group were higher than those of the middle-dose group and the high-dose group(P<0.05),so were in the medium dose group than those in the high dose group(P<0.05).Western blotting indicated that the relative expression levels of JAK2,STAT3 and Ki-67 proteins significantly differed from each other of the 4 groups(F=25.241,18.102,14.997,P<0.05).The levels of the 3 proteins were all decreased in the 3 treatment groups than the control group(P<0.05),and the levels of JAK2 and STAT3 were higher in the low-dose group were than the medium-dose group and the high-dose group(P<0.05),and also was higher in the medium-dose group than the high-dose group(P<0.05).Conclusion Saikosaponin D can inhibit the proliferation and promote the apoptosis in breast cancer MCF-7 cells,which may be closely related to the JAK2/STAT3 pathway.
作者
冯杰鑫
吴雄
陈建辉
林青海
叶泽霖
FENG Jiexin;WU Xiong;CHEN Jianhui;LIN Qinghai;YE Zelin(Department of BreastSurgery,Zhangzhou Hospital Affiliated to Fujian Medical University,Zhangzhou,Fujian Province,363000,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2021年第9期852-857,共6页
Journal of Third Military Medical University
