人参皂苷CK联合5-氟尿嘧啶对人胰腺癌PANC-1细胞的增殖、凋亡及上皮间质转化的影响

Effects of Ginsenoside CK Combined with 5-Fluorouracil on the Proliferation,Apoptosis and Epithelial Mesenchymal Transition of Human Pancreatic Cancer PANC-1 Cells

目的:研究人参皂苷CK联合5-氟尿嘧啶(5-FU)对人胰腺癌PANC-1细胞的增殖、凋亡及上皮间质转化(EMT)的影响。方法:将对数生长期的PANC-1细胞分为空白对照组、人参皂苷CK组(30 mg/L)、5-FU组(25 mg/L)和联用组(人参皂苷CK 30 mg/L+5-FU 25 mg/L)。采用MTT法检测各组细胞作用24、48、72 h后的细胞增殖抑制率,流式细胞术检测作用48 h后的细胞凋亡率,酶联免疫吸附法检测作用24、48、72、96 h后细胞中纤连蛋白表达,Western blot法检测作用48 h后细胞中波形蛋白、N-钙黏蛋白、E-钙黏蛋白、蛋白激酶(Akt)及磷酸化Akt(p-Akt)蛋白表达。结果:人参皂苷CK、5-FU及其二者联用对细胞的增殖均有抑制作用;与空白对照组比较,人参皂苷CK组、5-FU组和联用组各作用时间点的早期和晚期凋亡率、E-钙黏蛋白表达水平均明显升高(P<0.05),纤连蛋白、波形蛋白、N-钙黏蛋白表达水平和p-Akt/Akt水平均明显降低(P<0.05),其中联用组上述指标效果均优于人参皂苷CK组和5-FU组(P<0.05)。结论:人参皂苷CK和5-FU均可抑制PANC-1细胞增殖、诱导细胞凋亡、抑制EMT,该作用可能与抑制磷脂酰肌醇3-激酶/Akt通路有关;且二者联用效果更好。

OBJECTIVE：To study the effects of ginsenoside CK combined with 5-fluorouracil（5-FU） on the proliferation,apoptosis and epithelial mesenchymal transition（EMT）of human pancreatic cancer PANC-1 cells. METHODS：PANC-1 cells of logarithmic growth phase were randomly divided into blank control group,ginsenoside CK group（30 mg/L）,5-FU group（25 mg/L）and combination group（ginsenoside CK 30 mg/L＋5-FU 25 mg/L）. MTT method was used to detect the cell proliferation inhibition rate in each group after 24,48,72 h;flow cytometry was used to detect the cell apoptosis rate after 48 h;enzyme-linked immunosorbent assay was used to detect the fibronectin expression in cells after 24,48,72,96 h;and Western blot was used to detect the expressions of vimentin,N-cadherin,E-cadherin,protein kinase（Akt）and phosphorylated Akt（p-Akt）protein in cells after 48 h. RESULTS：Compared with blank control group,the cell proliferation inhibition rate,early and late apoptotic rates,protein expression level of E-cadherin in ginsenoside CK group,5-FU group and combination group were obviously increased（P〈0.05）,while the protein expression levels of fibronectin,vimentin,N-cadherin,and p-Akt/Akt levels were obviously decreased（P〈0.05）;the effects of above-mentioned indexes in combination group were superior to ginsenoside CK group and 5-FU group（P〈0.05）. CONCLUSIONS：Both ginsenoside CK and 5-FU can inhibit the proliferation of PANC-1 cells,induce their apoptosis and inhibit EMT,which may be associated with inhibiting phosphatidylinositol 3-kinase/Akt pathway. In addition,the combination of ginsenoside CK and 5-FU can produce a better effect.