肝胆胰外科杂志2025，Vol.37Issue(11)：764-771,776,9.DOI:10.11952/j.issn.1007-1954.2025.11.007
柴胡皂苷D通过丝裂原活化蛋白激酶及蛋白激酶B信号通路诱导胰腺癌细胞凋亡
Saikosaponin D induces apoptosis of pancreatic cancer cells through mitogen activated protein kinase and protein kinase B signaling pathways
摘要
Abstract
Objective This study aimed to investigate the apoptosis-inducing effects of Saikosaponin D(SSD),a bioactive compound derived from Bupleurum extract,on pancreatic cancer cells(Panc-1 cell line),and to preliminarily examine whether its mechanism involves the signaling pathways modulation of mitogen-activated protein kinase(MAPK)and protein kinase B(AKT).Methods Panc-1 cells were exposed to SSD at concentrations of 0(the control group),10 μmol/L and 20 μmol/L.Cell proliferation was evaluated using a colony formation assay.Apoptosis was detected by Annexin V-FITC/PI double staining followed by fluorescence microscopy and further quantified by flow cytometry.Cellular senescence was assessed via β-galactosidase staining under light microscopy.The mRNA expression levels of Caspase-3,-6,-7,-8,and-9 were determined using quantitative real-time PCR(qRT-PCR)after treatment with 20 μmol/L SSD.Protein levels of caspase-3,cleaved caspase-3,p38,phospho-p38(p-p38),AKT,and phospho-AKT(p-AKT)were analyzed by Western blotting(WB).Results Compared with the control group,SSD treatment(10 μmol/L and 20 μmol/L)significantly reduced the colony formation capacity(F=8.07,P<0.05).Apoptosis analysis revealed enhanced fluorescence signals and a notably increased total apoptosis rate—up to 79.14%—as measured by flow cytometry.β-Galactosidase staining showed elevated blue precipitate formation,indicating SSD-induced cellular senescence.qRT-PCR results indicated that 20 μmol/L SSD significantly upregulated the mRNA expression of all caspases tested(Caspase-3:t=-5.5713,P=0.0099;Caspase-6:t=-4.4721,P=0.0141;Caspase-7:t=-3.7639,P=0.0267;Caspase-8:t=-6.0000,P=0.0075;Caspase-9:t=-8.0000,P=0.0031).WB analysis demonstrated that SSD promoted the expression of cleaved Caspase-3 and p-p38,an effect partially abolished by the p38 inhibitor SB203580,implying involvement of the p38 MAPK pathway.Conversely,SSD also elevated p-AKT and cleaved Caspase-3 levels;The AKT agonist SC-79 partially suppressed cleaved Caspase-3 upregulation,suggesting an antagonistic role of AK activation against SSD-induced apoptosis.Conclusion This experiment suggests that Saikosaponin D can promote apoptosis in Panc-1 cell line,in which the MAPK and AKT signaling pathways are possibly involved.关键词
胰腺癌/柴胡皂苷D/Panc-1细胞/丝裂原活化蛋白激酶信号通路/蛋白激酶B信号通路/细胞凋亡Key words
pancreatic cancer/Saikosaponin D/Pan cell line/mitogen-activated protein kinase signalling pathway/protein kinase B signalling pathway/cell apoptosis分类
医药卫生引用本文复制引用
李仕珺,姚佳锜,王韬,吴斌,沈亦钰..柴胡皂苷D通过丝裂原活化蛋白激酶及蛋白激酶B信号通路诱导胰腺癌细胞凋亡[J].肝胆胰外科杂志,2025,37(11):764-771,776,9.基金项目
浙江省中医药管理局科研课题(2021ZB294). （2021ZB294）
