姜黄素联合吉西他滨对胰腺癌细胞体外增殖、凋亡及肿瘤干细胞表型特征的影响

摘要

目的：研究联合使用姜黄素及吉西他滨对胰腺癌细胞增殖、凋亡及肿瘤干细胞表型特征的影响，初步评价姜黄素治疗胰腺癌的临床应用前景。方法采用恶性程度、分化程度不同的胰腺癌细胞系panc03.27、Capan‐2及PANC‐1细胞，分为DM SO处理组、单纯吉西他滨处理组、单纯姜黄素处理组以及吉西他滨联合姜黄素处理组，采用CCK8（Cell Counting Kit‐8）试剂盒测定细胞增殖力的改变；同时采用FITC Annexin V凋亡检测试剂盒分析不同处理组之间细胞凋亡率的差异，利用Western blot检测胰腺癌干性基因Notch1及Oct4的表达改变。结果 Panc03.27细胞中与单纯使用吉西他滨比较，联合使用姜黄素组细胞活力由（46.70±3.49）％下降为（35.53±1.84）％（ P＝0.047），细胞凋亡率由（5.01±0.75）％上升至（13.11±0.89）％（P＜0.01），Notch1及Oct4表达明显下调（均 P＜0.01）；Capan‐2细胞中与单纯使用吉西他滨比较，联合使用姜黄素组细胞活力由（47.13±2.39）％下降至（5.36±0.25）％（ P＜0.01），细胞凋亡率由（13.02±1.79）％上升至（20.11±1.11）％（P＜0.01），而Notch1及Oct4表达未见明显差异；PANC‐1细胞中与单纯使用吉西他滨比联合使用姜黄素组细胞活力由（82.59±5.69）％下降至（7.33±0.25）％（ P＜0.01），细胞凋亡率由（6.77±2.03）％上升至（25.08±3.14）％（P＜0.01），Notch1及Oct4表达明显下调（P＜0.05）。结论姜黄素可协同吉西他滨抑制胰腺癌细胞的增殖、诱导细胞的凋亡及抑制肿瘤干细胞表型特征。%Objective To investigate the effect of combination therapy with curcumin and gemcitabine on the proliferation , apoptosis and cancer stem cell phenotype of pancreatic cancer cells in order to preliminarily examine the application potential of curcumin to the treatment of pancreatic cancer.Methods Human pancreatic cancer cell lines of different malignancy and differ‐entiation ,panc03.27 ,Capan‐2 and PANC‐1 ,were used in this study.They were divided into DMSO group ,gemcitabine treat‐ment group ,curcumin treatment group and gemcitabine+curcumin treatment group in terms of different treatments.Cell count‐ing Kit‐8 assay and flow cytometry were employed to detect the proliferation activity and apoptosis rate of these pancreatic canc‐er cells ,respectively.The protein expression of genes associated with cancer stem cells (Notch1 and Oct4)was measured by Western blot.Results Comparison of the proliferation and apoptosis between the single gemcitabine group and the combination group found that ,in panc03.27 cells ,the cell viability was reduced from(46.70 ± 3.49)% to(35.53 ± 1.84)% (P=0.047) ,the apoptosis rate was increased from(5.01 ± 0.75)% to(13.11 ± 0.89)% (P<0.01)and the expression levels of Notch1 and Oct4 were suppressed(P< 0.01);in Capan‐2 cells ,the cell viability was reduced from(47.13 ± 2.39)% to(5.36 ± 0.25)% (P<0.01) ,the apoptosis rate was increased from(13.02 ± 1.79)% to(20.11 ± 1.11)% (P< 0.01) ,and the expression levels of Notch1 and Oct4 were not significantly different;in PANC‐1 cells ,the cell viability was reduced from(82.59 ± 5.69)% to(7.33 ± 0.25)% (P<0.01) ,the apoptosis rate was increased from(6.77 ± 2.03)% to(25.08 ± 3.14)% (P<0.01) ,and the expres‐sion levels of Notch1 and Oct4 were suppressed(P<0.05).Conclusion Curcumin can synergistically inhibit the proliferation , promote the apoptosis of pancreatic cancer cells ,and suppress the phenotype of cancer stem cells when used with gemcitabine.