抑制Hedgehog信号通路改变结肠癌细胞基因表达谱  

Gene Expression Profiling of Hedgehog Signaling Pathway Inhibition in Human Colon Cancer Cells

作　　者：黄璇[1] 张伟龙[2] 罗清甜[1] 李勇[1] 张君严[1] 邓立彬[2] 吕农华[3] 罗时文[1] 

机构地区：[1]南昌大学第一附属医院医学科研中心,南昌330006 [2]南昌大学转化医学研究院,南昌330031 [3]南昌大学第一附属医院消化内科,南昌330006

出　　处：《中国细胞生物学学报》2012年第9期886-898,共13页Chinese Journal of Cell Biology

基　　金：国家重点基础研究发展计划(973计划)项目(No.2010CB535001);国家自然科学基金(No.81060095;No.31171359);江西省主要学科学术和技术带头人培养计划(No.2009DD00600);中国博士后基金(No.20100-481187);江西省自然科学基金(No.20114BAB215019);国家大学生创新性实验计划项目(No.091040305)资助项目~~

摘　　要：Hedgehog(Hh)信号通路在机体发育和肿瘤发生中发挥着重要作用。在该研究中,Western blot检测三株结肠癌细胞Hedgehog信号通路组分的表达,结果表明三株结肠癌细胞中HT-29细胞Hedgehog信号通路组分较完整。采用MTT和BrdU法检测Hedgehog信号通路膜受体Smo特异性抑制剂环杷明和末端转录因子Gli1/2的特异性抑制剂GANT61对HT-29细胞的影响,提示这两种抑制剂均显著抑制HT-29细胞生存率和细胞增殖率,且GANT61比环杷明更敏感。表达谱芯片检测阻断Hedgehog信号通路后HT-29细胞基因谱的变化,结合生物信息学分析,揭示HT-29细胞经环杷明和GANT61处理后基因表达呈现抑制特征,其差异基因表达主要以下调为主,其中环杷明主要影响细胞内源刺激等,而GANT61主要影响代谢和类固醇合成,并与MAPK信号通路有关联,两者均能影响细胞免疫及凋亡相关通路。这些结果提示,Hh信号通路有可能作为结肠癌的治疗靶点。Hedgehog signaling pathway plays an important role in animal development and tumorigenesis.In the present study,components of hedgehog signaling pathway were detected in HT-29 cells other than SW-480 or Caco-2 cells by Western blot.Blocking of hedgehog signaling by GANT61,which is a small molecule inhibitor of Gli1/2,can significantly decrease the survival rate and proliferation of colon cancer cells,whereas cyclopamine,the conventional Smo inhibitor,showed only modest cytotoxic activity.Gene expression profiling of hedgehog signaling inhibited HT-29 cells was determinated by cDNA microarray.Results indicated that gene expression was partially blocked and differential expressed genes were primarily knocked-down.Moreover,cyclopamine may be principally involved with cell endogenous stimulus,while GANT61 was related to metabolic and steroid biosynthesis and MAPK signaling pathway.Both of them affect pathways in celluar immunity and cell apoptosis.Our findings suggest that inactivation of hedgehog signaling may be an attractive target for the treatment of colon cancer.

关 键 词：HEDGEHOG信号通路 结肠癌 基因表达谱 

分 类 号：R735.35[医药卫生—肿瘤]