Identification of RegIV as a novel GLI1 target gene in human pancreatic cancer.
Identification of RegIV as a novel GLI1 target gene in human pancreatic cancer.
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鉴定 RegIV 作为人类胰腺癌的新型 GLI1 靶基因
DOI：
10.1371/journal.pone.0018434
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发表时间：
2011-04-11
期刊：
影响因子：
3.7
通讯作者：
Wang X
中科院分区：
文献类型：
--
作者：
Wang F;Xu L;Guo C;Ke A;Hu G;Xu X;Mo W;Yang L;Huang Y;He S;Wang X
背景与目的GLI 1是胰腺癌Hedgehog信号通路中的关键转录因子。RegIV与再生、细胞生长、存活、粘附和抗凋亡相关。我们的目的是研究RegIV在胰腺癌中的表达及其与GLI 1的关系。方法采用qRT-PCR、Western blot和免疫组化方法检测胰腺癌患者癌组织、癌旁正常组织和5株胰腺癌细胞系中GLI 1和RegIV的表达，并分析其相关性。构建GLI 1-shRNA慢病毒载体并转染PANC-1，构建含GLI 1表达序列的慢病毒载体并转染BxPC-3。通过qRT-PCR和Western印迹评价GLI 1和RegIV表达。最后，我们通过染色质免疫沉淀（CHIP）和电泳迁移率变动分析（EMSA）证明RegIV是GLI 1的靶点。结果免疫组化和qRT-PCR结果显示，胰腺癌组织中RegIV和GLI 1的表达均高于癌旁正常组织（p<0.001）。在这些组织中，RegIV表达与GLI 1表达相关（R = 0.795，p<0.0001）。 这些结果在5种胰腺癌细胞系中的蛋白质（R= 0.939，p = 0.018）和mRNA表达（R = 0.959，p =0.011）得到验证。 当用RNAi技术敲低GLI 1时（分别为82.1± 3.2%，76.7±2.2%;），RegIV mRNA和蛋白表达均降低（分别为94.7± 0.3%，84.1±0.5%;）。GLI 1在mRNA和蛋白水平的过表达（分别为924.5± 5.3%和362.1±3.5%）诱导RegIV的过表达（分别为729.1± 4.3%和339.0±3.7%）。此外，CHIP和EMSA测定显示GLI 1蛋白结合到胰腺癌细胞中的RegIV启动子区域（GATCATCCA）。结论胰腺癌中GLI 1通过与RegIV基因启动子结合促进RegIV的转录。
Background and Aims GLI1 is the key transcriptional factor in the Hedgehog signaling pathway in pancreatic cancer. RegIV is associated with regeneration, and cell growth, survival, adhesion and resistance to apoptosis. We aimed to study RegIV expression in pancreatic cancer and its relationship to GLI1. Methods GLI1 and RegIV expression were evaluated in tumor tissue and adjacent normal tissues of pancreatic cancer patients and 5 pancreatic cancer cell lines by qRT-PCR, Western blot, and immunohistochemistry (IHC), and the correlation between them. The GLI1-shRNA lentiviral vector was constructed and transfected into PANC-1, and lentiviral vector containing the GLI1 expression sequence was constructed and transfected into BxPC-3. GLI1 and RegIV expression were evaluated by qRT-PCR and Western blot. Finally we demonstrated RegIV to be the target of GLI1 by chromatin immunoprecipitation (CHIP) and electrophoretic mobility shift assays (EMSA). Results The results of IHC and qRT-PCR showed that RegIV and GLI1 expression was higher in pancreatic cancer tissues versus adjacent normal tissues (p<0.001). RegIV expression correlated with GLI1 expression in these tissues (R = 0.795, p<0.0001). These results were verified for protein (R = 0.939, p = 0.018) and mRNA expression (R = 0.959, p = 0.011) in 5 pancreatic cancer cell lines. RegIV mRNA and protein expression was decreased (94.7±0.3%, 84.1±0.5%; respectively) when GLI1 was knocked down (82.1±3.2%, 76.7±2.2%; respectively) by the RNAi technique. GLI1 overexpression in mRNA and protein level (924.5±5.3%, 362.1±3.5%; respectively) induced RegIV overexpression (729.1±4.3%, 339.0±3.7%; respectively). Moreover, CHIP and EMSA assays showed GLI1 protein bound to RegIV promotor regions (GATCATCCA) in pancreatic cancer cells. Conclusion GLI1 promotes RegIV transcription by binding to the RegIV gene promoter in pancreatic cancer.
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影响因子：
29.4
作者：
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通讯作者：
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作者：
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