雷公藤内酯醇联合吉西他滨对胰腺癌细胞增殖和凋亡的影响

Effect of triptolide combined with gemcitabine on proliferation and apoptosis of pancreatic cancer cells

目的观察雷公藤内酯醇(TPL)联合吉西他滨(GEM)对胰腺癌PANC-1细胞增殖和凋亡的影响。方法以TPL、GEM单独用药以及两药联合作用于体外培养的PANC-1细胞,采用细胞毒实验(MTT)法检测细胞增殖并计算两药之间的交互作用、Hoechst33258染色观察细胞凋亡形态学变化、流式细胞仪检测细胞凋亡率、蛋白免疫印迹法(Western blot)检测信号转导与转录活化因子3(STAT3)、半胱氨酸天冬氨酸特异性蛋白酶-3(caspase-3)蛋白表达。结果 TPL、GEM单药以及联合用药组的细胞均出现了典型的细胞凋亡形态学改变,且联合用药组的凋亡细胞数较TPL、GEM单药组明显增加。与对照组相比,TPL、GEM单药以及联合用药组的细胞凋亡率均明显增加,差异有统计学意义(P<0.05);且联合用药组的细胞凋亡率显著高于单药组,差异有统计学意义(P<0.05)。TPL联合GEM可协同抑制兔抗人磷酸化转录活化因子3(p-STAT3)蛋白表达,活化caspase-3TPL、GEM蛋白表达。结论 TPL联合GEM可协同抑制胰腺癌PANC-1细胞增殖并诱导细胞凋亡,其机制与抑制STAT3通路、促进caspase-3蛋白表达有关。

Objective To observe the effect of triptolide combined with gemcitabine on proliferation and apoptosis of pancreatic cancer cells ,and to analyze the relevant mechanisms .Methods After treated with TPL ,GEM or TPL combined with GEM in vitro , PANC-1 cells proliferation was accessed by MTT assay and the interaction between the two drugs was calculated .Apoptotic mor-phological changes and apoptosis rate of the cells were investigated by Hoechst 33258 staining and flow cytometry ,respectively .The expression of signal transduction and transcription factor 3（STAT3） ,cysteine aspartate specific proteases-3（caspase-3） protein were detected by Western blot analysis .Results TPL ,GEM or TPL combined with GEM could significantly inhibit the prolifera-tion of PANC-1 cells ,and the combination of the two drugs had a synergistic effect .The cells of the TPL group ,GEM group ,as well as the combined group showed typical apoptotic morphological changes .Compared with the TPL group and GEM group ,the number of apoptotic cells of the combined group increased significantly .Compared with the control group ,the cells apoptosis rate of the TPL group ,GEM group and combined group was significantly increased （P〈 0 .05） ,and the apoptosis rate of the the combined group was significantly higher than that of the monotherapy group（P〈0 .05） .TPL combined with GEM synergistically inhibited p-STAT3 protein expression and activated caspase-3 protein expression .Conclusion TPL combined with GEM can synergistically in-hibit proliferation and induce apoptosis of pancreatic cancer PANC-1 cells ,its mechanism is related to the inhibition of STAT 3 sig-naling pathway ,promotion the expression of caspase-3 protein .