右旋柠烯对HL60K562白血病细胞体外作用的研究.docVIP

右旋柠烯对HL60K562白血病细胞体外作用的研究 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：右旋柠烯对HL60K562白血病细胞体外作用的研究 1 文2：PI103对多药耐药白血病细胞体外作用的研究 8 1 材料和方法 9 2 结果 10 3 讨论 11 参考文摘引言： 12 原创性声明（模板） 14 文章致谢（模板） 14 正文 右旋柠烯对HL60K562白血病细胞体外作用的研究 文1：右旋柠烯对HL60K562白血病细胞体外作用的研究 Effects of D-limonene on Leukemia Cells HL-60 and K562 in Vitro Abstract To investigate the effects of D-limonene(D-L) on the cell growth and apoptosis in HL-60，K562 cells and to elucidate its mechanism，the influence of D-L on proliferation of HL-60 and K562 cells was determined by propidium iodide assay，the expression levels of mutant p53，bcl-2，bax gene were detected by cell morphological analysis，flow cytometry and immunohistochemistry staining，the D-L-inducing HL-60 and K562 cell apoptosis in vitro was observed systematically. The results showed that D-L inhibited HL-60 and K562 cell growth in a dose-and time-dependent manner with the IC50 of mmol/L similarly，D-L induced apoptosis of HL-60 and K562 cells，and expression of bcl-2 gene was down regulated by D-L in a concentration-dependent manner in HL-60 bcl-2，mutant type of p53 genes were down regulated while bax gene was up regulated by D-L in a concentration-dependent manner in K562 cells. It is concluded that D-L can inhibit proliferation and induce apoptosis of HL-60 and K562 cells. The bcl-2，mutant type of p53 and bax may be involved in the gene regulation of D-L-induced apoptosis. Key words D-limonene; HL-60，K562; Cell proferation; Apoptosis 右旋柠烯（D－limonene，D-L）是一种天然单萜烯类化合物，在桔皮精油中的含量90%，学名1-甲基-4-异丙基环己烯，分子式C10H16，分子量。D-L是limonene中最普遍存在的一种异构体，具有新鲜柠檬香气和抗氧化、抗炎、利胆溶石等作用。近年发现D-L具有抗癌活性，用于预防及 治疗 化学致癌物诱导的结肠癌、乳腺癌、胃癌、肺癌、皮肤癌、肝癌、胰腺癌等取得了相当的效果［1-3］。目前对其抗癌作用的研究主要集中其对实体瘤的作用机制上，关于其对非实体瘤的作用的研究报道尚少。本实验选用HL-60、K562白血病细胞，体外观察D-L对这些细胞增殖抑制及诱导凋亡的影响，并进一步探讨其作用机制，以期寻找新的安全有效的预防和治疗白血病的制剂。 材料和方法 材料 右旋柠烯购自美国Sigma公司，含量97%，用二甲亚砜(DMSO)稀释至所需浓度。DMEM培养液(含10%胎牛血清、100 U/ml青霉素和100 μg/ml链霉素，pH )，碘化丙锭(propidium iodide，PI )，毛地黄皂苷（digitonin)，DMSO均购自于Sigma公司。突变型p53，bcl-2及bax鼠抗人即用型IgG单克隆抗