二氢青蒿素对胰腺癌生长的抑制作用

Study on anticancer effect of dihydroartemisinin on pancreatic cancer

目的探讨二氢青蒿素在体外、体内对胰腺癌的生长抑制作用。方法通过MTF法检测二氢青蒿素对胰腺癌细胞株生长的抑制作用，Annexin V—FITC／PI染色流式细胞术检测细胞凋亡；Western blot检测BxPC-3细胞中增殖、凋亡相关蛋白的表达。监测给药后胰腺癌裸鼠移植瘤体积的变化，并通过对肿瘤组织标本Ki-67染色和TUNEL染色检测肿瘤细胞增殖和凋亡情况。结果MTT结果显示，二氢青蒿素可抑制体外培养的胰腺癌细胞BxPC-3和ASPC-1的增殖，诱导细胞凋亡，且呈剂量依赖性。二氢青蒿素亦可通过抑制增殖和诱导凋亡而抑制胰腺癌的体内生长。Western blot检测BxPC-3细胞中蛋白的表达水平，结果显示二氢青蒿素上调增殖相关蛋白p21^WAF1、下调PCNA的表达；上调凋亡相关蛋白Bax、下调Bcl-2的表达，且可增加caspase-9的活化水平。结论二氢青蒿素在体内外对胰腺癌均有抗肿瘤作用，是胰腺癌治疗的潜在药物。

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITC/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to （ 76. 2±3.5 ） % and （ 79.5±2. 9） % , and the apoptosis rates were up to （55. 5±2）% and （40. 0±3.5）%, the differences were significantly （ P 〈 0. 01 ） compared with control [ （2. 0±1.3 ） % and （0.9 ±0.4） %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptosis index were （49. 1±3.9） % and （ 50. 2±4.4 ） % respectively in dihydroartemisinin 50 mg/kg treatment group, compared to those of （ 72.1±3.3 ） % and （ 9. 4±2. 9 ） % in control, the differences were significantly （ P 〈 0. 01 ）. Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferationassociated protein p21^WAF1 and down-regulates expression of PCNA, increases expression of apoptosisassociated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptosis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.