青蒿琥酯增强吉西他滨的抗胰腺癌活性

Artesunate Enhances the Anti-tumor Effect of Gemcitabine on Pancreatic Cancer

关键词:  青蒿琥酯  FOXO1  吉西他滨  胰腺癌

Key words:  artesunate  FOXO1  gemcitabine  pancreatic cancer

摘要:

目的 探讨青蒿琥酯对吉西他滨化疗胰腺癌的辅助治疗作用并研究其机制。方法 MTT法检测胰腺癌细胞系Capan-2在青蒿琥酯和不同浓度吉西他滨处理下的细胞活力。Western blot检测吉西他滨和青蒿琥酯对Capan-2细胞FOXO1、Noxa、Bim表达水平，细胞色素C、Smac/DIABLO从线粒体中的释放量及caspase-9、caspase-3活化水平的影响。流式细胞术检测Capan-2细胞的线粒体膜电位和凋亡率。结果 青蒿琥酯辅助治疗明显提高吉西他滨对Capan-2细胞的杀伤活性。青蒿琥酯处理能显著促进Capan-2细胞FOXO1的表达，转染FOXO1小干扰RNA （FOXO1 siRNA）后，青蒿琥酯对吉西他滨的辅助治疗效果受到明显抑制。青蒿琥酯联合吉西他滨能显著诱导Capan-2细胞中Noxa和Bim的过表达，线粒体膜电位的降低，细胞色素C、Smac/DIABLO的释放，caspase-9、caspase-3的活化及凋亡的发生。转染FOXO1siRNA后，青蒿琥酯联合吉西他滨对Capan-2细胞的凋亡诱导途径受到显著抑制。结论 青蒿琥酯可上调FOXO1的表达，增强吉西他滨对胰腺癌细胞的凋亡诱导活性。

Abstract:

OBJECTIVE To investigate the adjuvant effect and mechanisms of artesunate on gemcitabine-based chemotherapy in pancreatic cancer. METHODS MTT assay was performed to evaluate the effect of artesunate and different concentrations of gemcitabine on changing the cell viability of Capan-2. Western blot analysis was performed to detect the effect of artesunate and gemcitabine on changing the expression of FOXO1, Noxa, Bim, and release of cytochrome C and Smac/DIABLO from mitochondria and activation of caspase-9, caspase-3 in Capan-2 cells. Flow cytometry analysis was performed to detect the mitochondrial membrane potential(Δφ) and apoptosis of Capan-2. RESULTS Adjuvant therapy of artesunate significantly enhanced the killing activity of gemcitabine against pancreatic cancer in vitro. Artesunate significantly promoted the expression of FOXO1 in Capan-2. However, transfection with FOXO1 siRNA obviously suppressed the adjuvant effect of artesunate on gemcitabine. Combination with gemcitabine and artesunate significantly induced the overexpression of Noxa and Bim, release of cytochrome C and Smac/DIABLO, activation of caspase-9 and caspase-3, suppression of Δφ and occurrence of cell apoptosis in Capan-2 cells. On the other hand, transfection with FOXO1 siRNA obviously suppressed the apoptotic pathway induced by the combination treatment with artesunate and gemcitabine in the Capan-2 cells. CONCLUSION Artesunate enhances the activity of gemcitabine on inducing the apoptosis of pancreatic cancer through the upregulation of FOXO1.