《Journal of Oral and Maxillofacial Surgery》 ›› 2012, Vol. 22 ›› Issue (3): 163-167. doi: 10.3969/j.issn.1005-4979.2012.03.003

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Cyclopamine Induces Apoptosis in Human Salivary Pleomorphic Adenoma Cells

OU Xue-ping1, XIAO Cui-cui1, SONG Meng1, XIAO Zhong-ming2, WEI Fang2, HONG Yan2   

  1. 1.Department of Stomatology, 2.Experimental Research Center, Shanghai Municipal First People’s Hospital, Shanghai Jiaotong University, Shanghai 200080, China
  • Online:2012-06-28 Published:2012-08-30

环杷明对人腮腺多形性腺瘤细胞凋亡的影响

欧学平1,肖翠翠1,宋萌1,肖忠明2,韦芳2,洪艳2   

  1. 1.上海交通大学附属上海第一人民医院口腔科,2.实验中心, 上海 200080
  • 通讯作者: 宋萌,教授. E-mail:smgjzh160@sohu.com
  • 作者简介:欧学平(1981—),男,湖北黄冈市人,硕士研究生. E-mail:omigaso@126.com
  • 基金资助:

     上海市科学技术委员会资助项目(08JC1412900);上海   市重点学科建设项目(S30206-KF05)

Abstract: Objective: The purpose of this study was to examine whether  cyclopamine is related to  apoptosis and growth inhibition of human salivary pleomorphic adenoma (HSPA) cells.  Methods: HSPA cells were treated  with cyclopamine.Observed the cells quantity change and morphology after 48 h. Cells cultured with DMSO 0.08% or void were used as control. Gli-2 and Bcl-2 mRNA expression was examined by Real-time PCR after 48 h. Flow cytometry was used to analyze cell apoptosis after 24 h. Experiment results are presented as x±s. Statistical analysis was undertaken with SAS 9.13. The differences among the groups were identified by one-way ANOVA. Significance was accepted at P<0.01. Result: The number of cyclopamine treated  HSPA cells was significantly decreased. Cyclopamine treated (10 μmol/L) HSPA cells showed  nuclear psychosis or fragmentation, chromatospherite disfiguration with apoptoticmorphology.  The expression levels of Gli-2 and Bcl-2 mRNA in 10 μmol∕L cyclopamine treated group was down-regulated significantly compare with blank and DMSO control groups (P<0.01). Apoptosis rate of 10 μmol∕L cyclopamine treatment group was significantly higher than blank and DMSO control group (P<0.01). Conclusion: Cyclopamine appears to induce growth inhibition and apoptosis in human salivary pleomorphic adenoma cells. The mechanism may be related to down-regulation of  Gli-2 and Bcl-2 mRNA, which activates the mitochondrial apoptotic pathways.

Key words: pleomorphic adenoma cell, apoptosis, cyclopamine

摘要:  目的:探讨环杷明(cyclopamine)诱导人腮腺多形性腺瘤细胞凋亡的作用及其对Gli-2和Bcl-2表达的影响。方法:用10 μmol∕L环杷明处理人腮腺多形性腺瘤细胞,48 h后倒置显微镜下观察细胞数量及形态变化;48 h后实时定量PCR检测实验组(环杷明组)和空白对照组、DMSO对照组的Gli-2、Bcl-2 mRNA表达水平差异;24 h后流式细胞术观察细胞凋亡。结果:人腮腺多形性腺瘤细胞用10 μmol∕L 环杷明作用48 h后倒置显微镜下可见细胞数量减少、细胞核固缩或碎裂、核仁变形等典型的细胞凋亡形态学变化;环杷明组与空白对照组、DMSO对照组相比,可以显著下调Gli-2、Bcl-2 mRNA表达(P<0.01);环杷明组的细胞凋亡率明显高于空白对照组和DMSO对照组(P<0.01)。结论:环杷明可诱导人腮腺多形性腺瘤细胞凋亡,其作用机制可能与下调Gli-2、Bcl-2 mRNA表达,活化细胞凋亡的线粒体途径有关。

关键词: 腮腺多形性腺瘤细胞, 细胞凋亡, 环杷明

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