牛蒡子苷对胰腺癌细胞抑制作用及其作用机理实验探究.doc

牛蒡子苷对胰腺癌细胞抑制作用及其作用机理实验探究 　【摘要】 目的通过牛蒡子苷对胰腺癌细胞株(CAPAN-1)体内外实验，验证其是否具有抗胰腺癌作用。方法在CAPAN-1细胞培养液中加牛蒡子苷，培养72 h并观察癌细胞的增殖抑制率及癌细胞形态变化。流式细胞仪（FCM）检测CAPAN-1细胞周期及凋亡率；TUNEL法检测CAPAN-1细胞凋亡率；免疫细胞化学检测CAPAN-1细胞的增殖凋亡调控相关蛋白(bax、bcl-2)表达。用CAPAN-1细胞制作Balb/c裸小鼠荷瘤模型，连续每天腹腔注射牛蒡子苷（10 mg·kg-1·d-1）10d，以5-FU（30 mg·kg-1·d-1）作阳性对照观察抑瘤率。结果牛蒡子苷抑制CAPAN-1细胞生长；阻滞CAPAN-1细胞周期于G0/G1期并诱导细胞凋亡；通过上调bax蛋白表达（P＜0.01），下调bcl-2蛋白表达（P＜0.05）诱导CAPAN-1细胞凋亡；抑制荷瘤裸小鼠肿瘤生长，抑瘤率为41.7%。结论牛蒡子苷具有抗胰腺癌活性，其作用机理之一是诱导细胞凋亡。 【关键词】 牛蒡子苷； 抗肿瘤活性； 细胞凋亡； CAPAN-1细胞株 Abstract：ObjectiveTo evaluate the anti-tumor effect and mechanism of arctiin on the human pancreas cancer cell (CAPAN-1) in vitro and vivo. MethodsCAPAN-1 cell was cultured in medium containing serial concentration of arctiin for 72 hours. Then the growth rate and the morphological change were analyzed. Cell apoptosis percentage was measured by TUNEL method and FCM. Cell cycle phase distribution was measured by FCM. The expression of proliferation and apoptosis associated proteins of bax,bcl-2 in CAPAN-1 cell were determined using immunocytochemistry. CAPAN-1 cell tumor-bearing nude mice were intraperitoneally administered arctiin and 5-FU and the tumor growth rate was observed.ResultsArctiin could inhibit proliferation of CAPAN-1 cell. The cell cycle was arrested in G0/G1 phase and apoptosis was induced（P＜0.05）. The onset of apoptosis was related to the up-regulation of bax protein（P＜0.01）and down-regulation of bcl-2 in protein level（P＜0.05）. Arctiin could inhibit the tumor growth, and inhibition rate was 41.7%.ConclusionThese studies suggest that arctiin can inhibit CAPAN-1 cells growth and one of mechnnisnms is to induce cell apoptosis. Key words：Arctiin； Anti-tumor effect； Apoptosis； CAPAN-1 cell line 牛蒡子是具有疏散风热、宣肺透疹、解毒利咽的中药，且常用于抗癌方剂中。牛蒡子主要含牛蒡子苷（arctiin）、牛蒡子苷元（arctigenin）、牛蒡子酚A、B（lappaol A,B）、脂肪油等［1］。近来研究表明，牛蒡子苷及牛蒡子苷元在动物体内实验中具有抗癌活性 ［2～4］。牛蒡子苷对CAPAN-1细胞是否具有抑制作用尚无报道，为进一步研究牛蒡子苷的抗肿瘤活性，利用CAPAN-1细胞进行体内外实验，探讨牛蒡子苷是否具有抗胰腺癌活性及其作用机理。 1 材料 牛蒡子苷购于中国药品生物制品检定所；CAPAN-1细胞株由上海细胞研究所提供；FCS-