去甲斑蝥素对胆管癌rbe 细胞α-tubulin 骨架蛋白及细胞周期的影响.pdfVIP

中华临床医师杂志( 电子版)2013 年11 月第7 卷第21 期 Chin J Clinicians(Electronic Edition),November 1,2013,Vol.7,No.21 ·9581 · ·基础论著· 去甲斑蝥素对胆管癌RBE 细胞α-Tubulin 骨架蛋白及细胞周期的影响 王天阳 王文斌 边伟 张小艳 刘建华 王凤安 【摘要】 目的 观察去甲斑蝥素对人胆管癌RBE 细胞周期的影响及对微管蛋白（α-Tubulin ）的作用。 方法 体外培养胆管癌RBE 细胞，分别用不同剂量去甲斑蝥素处理，流式细胞学检测细胞周期的影响， 免疫组化染色法观察α-Tubulin 结构变化，Real-time PCR 检测α-Tubulin mRNA 转录水平变化。结果 流 式细胞学检测显示胆管癌RBE 细胞随着药物浓度的增加，G2/M 期阻滞作用增强，10、20 、40 μg/ml 组与 对照组相比具有统计学意义（P ＝0.000，P ＜0.01 ）。免疫组化染色法观察到去甲斑蝥素作用下，胆管癌RBE 细胞α-Tubulin 骨架结构被破坏，Real-time PCR 证实去甲斑蝥素使胆管癌RBE 细胞α-Tubulin 转录水平减 少，差异具有统计学差异（P ＝0.002，P ＝0.000，P ＜0.01 ）。结论 去甲斑蝥素可以通过破坏胆管癌RBE 细胞Tubulin 骨架、抑制α-Tubulin mRNA 的表达，阻止纺锤体的形成，将细胞周期阻滞在G2/M 期，导致 生长抑制，是其抗肿瘤机制之一。 【关键词】 胆管肿瘤； 细胞骨架； 细胞周期； 微管蛋白； 去甲斑蝥素 Norcantharidin induces G2/M cell cycle arrest though disrupting α-Tubulin cytoskeleton WANG Tian-yang, WANG Wen-bin, BIAN Wei, ZHANG Xiao-yan, LIU Jian -hua, WANG Feng-an. Department of Hepatobiliary Surgery, the Second Hospital, Hebei Medical University, Shijiazhuang 050000, China Corresponding author: LIU Jian-hua, Email: ljh@ 【Abstract 】 Objective To explore the mechanisms of Norcantharidins anti-tumor effect by investigating cell cycle and α-Tubulin cytoskeleton changes in cholangiocarcinoma cell line RBE after treated by Norcantharidin. Methods The RBE cells were cultured in vitro, followed by treating with various concentration of norcantharidin for 24 h. Cell cycle changes were detected by flow cytometric analysis. We used immunohistochemical (IHC) to observe α-Tubulin cytoskeleton changes, and real-time PCR was used to detect α-Tubulin mRNA transcript levels. Results The result of flow cytometric analysis shown the cell cycle arrested at G2/M. The damages of α-Tubulin cytoskeleton under the treatment of Norcantharidin were observed by IHC, and real-time PC