白藜芦醇和吉西他滨联合用药对人胰腺癌细胞作用的研究

Study on Synergistic Effect of Resveratrol and Gemcitabine on Human Pancreatic Cancer Cells

背景:吉西他滨是治疗进展期胰腺癌的一线化疗药物,单独用药临床效果欠佳。白藜芦醇为脱嘌呤脱嘧啶核酸内切酶1/氧化还原因子-1(APE1/Ref-1)的氧化还原功能抑制剂,具有抑制恶性肿瘤生长的特性,可能在胰腺癌的预防和治疗中发挥重要作用。目的:检测白藜芦醇和吉西他滨联合用药对人胰腺癌细胞株生长和凋亡的影响,并进一步探讨发挥该作用的分子机制。方法:将胰腺癌细胞株SW1990和BxPc-3分为4组:溶剂对照组、吉西他滨组、白藜芦醇组和联合用药组。采用CCK-8法和流式细胞术分别检测细胞增殖和凋亡,以蛋白质印迹法检测APE1/Ref-1蛋白的表达。结果:作用于SW1990和BxPc-3细胞24 h、48 h和72 h后,与溶剂对照组相比,三组用药组的细胞存活率均明显降低;作用于SW1990和BxPc-3细胞48 h后,与溶剂对照组相比,三组用药组的细胞凋亡率均明显增加;联合用药组对细胞增殖和凋亡的影响均强于两组单独用药组。与空白对照组相比,吉西他滨组SW1990和BxPc-3细胞APE1/Ref-1蛋白表达均明显增加。结论:白藜芦醇和吉西他滨联合用药可明显加强对胰腺癌细胞增殖和凋亡的影响,白藜芦醇可能通过抑制APE1/Ref-1蛋白的氧化还原功能而增加胰腺癌细胞对吉西他滨的敏感性。

Background： Gemcitabine is a first line chemotherapeutic drug for the treatment of advanced pancreatic cancer and in clinical practice it is not very effective when used alone. Resveratrol is an apurinic/apyrimidinic endonuclease 1/redox factor-1 （APEI/Ref-1） redox inhibitor, inhibiting the growth of malignant neoplasms and having a promising role in the prevention and treatment of pancreatic cancer. Aims： To examine the synergistic effect of resveratrol and gemcitabine on growth and apoptosis of human pancreatic cancer ceils, and to explore the possible molecular mechanism. Methods： Pancreatic cancer cell lines SW1990 and BxPc-3 were divided into 4 groups： vehicle control group, gemcitabine group, resveratrol group and gemcitabine combined with resveratrol group. Cell proliferation and apoptosis were determined by CCK-8 assay and flow cytometry, respectively; expression of APE1/Ref-1 protein was detected by Western blotting. Results： After intervention for 24, 48, 72 hours, proliferation rates of SW1990 and BxPc-3 cells in gemcitabine group, resveratrol group and gemcitabine combined with resveratrol group were significantly decreased when compared with vehicle control group. After intervention for 48 hours, apoptosis rates of SW1990 and BxPc-3 cells in the three intervention groups were significantly higher than those in vehicle control group. The effects on proliferation and apoptosis in gemcitabine combined with resveratrol group were significantly higher than either gemcitabine or resveratrol alone group. Compared with blank control group, expression of APE1/Ref-1 protein in SW1990 and BxPc-3 cells in gemcitabine group was significantly increased. Conclusions： The inhibition of cell growth and induction of apoptosis are significantly higher by the combination of resveratrol with gemcitabine. Resveratrol may significantly sensitize pancreatic cancer cells to the effect of gemcitabine via inhibiting the oxidation-reduction function of APE1/Ref-1 protein.