【作者】 彭华毅；

【导师】 林菁；

【作者基本信息】 福建医科大学 ， 药理学， 2010， 硕士

【摘要】 目的:探讨黄癸素的体内外抗肿瘤作用及其诱导细胞凋亡、分化等抗肿瘤作用机制。方法:⑴应用MTT法检测黄癸素对体外培养的HepG2、MGC80-3、SW480等13株肿瘤细胞增殖的抑制作用。⑵应用小鼠移植性实体瘤H22、S180和裸鼠移植瘤Colon26的治疗试验观察黄癸素的体内抗肿瘤活性。⑶采用吸光度法观察黄癸素对B16细胞黑色素含量的影响,探讨其诱导细胞分化作用。⑷通过Rhodamine 123染色、琼脂糖凝胶电泳、Hoechst 33258染色、caspase3和caspase 8活性检测,观察黄癸素诱导细胞凋亡作用。⑸采用免疫组化法检测裸鼠Colon26移植瘤CD44V6的表达,并观察黄癸素的影响。结果:黄癸素显著抑制体外培养HepG2、MGC80-3、SW480等肿瘤细胞的增殖;显著抑制小鼠H22、S180实体瘤的在体生长,对裸鼠移植瘤Colon26也有明显抑制作用;经黄癸素处理后的B16细胞黑色素生成能力增强,细胞趋向分化成熟;经黄癸素处理后,Rhodamine 123的荧光强度明显降低,琼脂糖凝胶电泳观察到DNA Ladder的出现,Hoechst 33258染色可观察到较高比例凋亡细胞,caspase3、caspase 8活性增强,显示黄癸素有促凋亡作用;黄癸素给药组Colon26移植瘤CD44V6表达较对照组降低,提示有抗肿瘤转移的可能。结论:黄癸素有较强的体内外抗肿瘤活性,并能诱导B16细胞分化、凋亡,降低CD44V6表达。更多还原

【Abstract】 Objective: To investigate the antitumor effects of berberineα-hydroxyβ-decanoylethyl sulfonate (HB), and its mechanism such as induction of cell apoptosis and differentiation.Methods: 1) MTT assay was employed to determine the cytotoxicity of HB in HepG2,MGC80-3, SW480 and other cells. 2) HB was administrated to treat mouse transplanted solid tumors H22 and S180,Colon26(a human cancer)in nude mice, and the tumor growth was observed. 3) The difference of melanin content in B16 cells between HB treatment group and control group was determined by using absorption photometry. 4)Through Rhodamine 123 dyeing, agarose gel electrophoresis, Hoechst 33258 dyeing, active detection of caspase 3 and caspase 8, observe the cell apoptosis induced by HB. 5)CD44V6 expression of Colon26 tumors which was transplanted in nude mice was detected by immunohistochemical technigue, and the influence of HB was observed on it.RESULT: HB significantly inhibits the proliferation of HepG2, MGC80-3, SW480 and other cells in vitro;inhibits the growth of H22 and S180 in mice, Colon26 tumors in nude mice. Melanin content of B16 cells increase after treatment with HB. The evidences of B16 cells apoptosis induced by HB were provided, such as Rhodamine 123 fluorescence intensity obviously decreased, DNA Ladder detected, nuclei pyknosed, caspase3 and caspase 8 activity increased. CD44V6 expression was decreased in Colon26 tumors which was treated with HB.Conclusion: HB has significant antitumor activity in vitro & in vivo, and can induce differentiation and apoptosis of B16 cells, reduce CD44V6 expression in Colon26 tumors.更多还原