抑制人肿瘤细胞Hedgehog 信号通路的基因表达谱分析
汤晓丽1，邓立彬2，张伟龙2，高猛3，黄登亮3，罗时文3，卢曲琴4

（1. 南昌大学生理教研室，南昌 330001；2. 南昌大学转化医学院，南昌 330001；3. 南昌大学第一附属医院医学科研中心，南昌 330006； 4. 南昌大学公卫学院，南昌 330006）
摘要：背景：在人类肿瘤中发现Hedgehog（Hh）信号通路异常激活，并且越来越多的证据表明GANT61（Gli 拮抗剂，是一种Hh 信号通路抑制剂），在抗癌治疗中起着相当大的作用。然而，目前仍然缺乏对Hh 抑制剂作用各种癌细胞的共同机制的系统扫描。方法/主要发现：Illumina® Sentrix® Bead 芯片分析出了Hh 抑制HT-29 和MKN45 的基因表达谱。17329个表达基因中，我们鉴定了在HT-29 和MKN45 细胞中分别有668 和269 个不同的基因表达，这两者之间具有差异表达（P<0.01）。发现大量的常见差异表达基因（77 个基因）均出现在这两株细胞中，这超出了预计数（10，P<0.0001）。基因本体论的进一步解释是基于过代表性分析。在这两株细胞中以基因本体生物过程的两个分类（“细胞死亡”和“对刺激物的响应”）筛选差异表达的基因。此外，对GANT61 处理的ES2 和H4 进行基因芯片分析，证实六个（CDKN1A， DDIT3，IER3，IL8，MFGE8 和PPP1R15A ）与“细胞死亡”相关基因差异表达。结论/意义：我们的研究表明，在各种肿瘤细胞中，抑制Hh 信号通路后，“细胞死亡”相关的基因有影响。并且，“DNA 损伤反应”相关的基因（如DDIT3）异常表达，在GANT61 诱导的细胞死亡中起着重要的作用。总之，这些数据有助于理解GNAT61 抗癌治疗活性的分子机制，以及在肿瘤细胞中发现GLI 新的靶基因。
 关键词：Hedgehog 信号通路; GANT61; 基因差异表达；抗癌治疗
中图分类号：R34

Gene Expression Profile of Hedgehog Signaling Pathway
Inhibition in Human Carcinoma Cells
TANG Xiaoli1, DENG Libin2, ZHANG Weilong2, GAO Meng3, HUANG Dengliang3,LUO Shiwen3, LU Quqin4**
(1. School of Medicine, Nanchang University, Nanchang, Jiangxi, 330001, China;2. Institute of Translational Medicine, Nanchang University, Nanchang, Jiangxi,330001, China;3. Center for Experimental Medicine, the First Affiliated Hospital of Nanchang University,Nanchang, Jiangxi, 330006, China;4. School of Public Health, Nanchang University, Nanchang, Jiangxi 330006, China)
Brief author introduction:TANG Xiaoli：Female,PhD Candiate,Lecturer,Research direction: molecular mechanisms
of diabetes Corespondance author: LU Quqin(1963-),Female,Associate Professor,Main research:System biology.

Abstract: Background: Aberrant activation of the Hedgehog (Hh) signaling pathway frequency occurs

 in human cancers, and increased evidence implicates the considerable role of GANT61
(Gli-ANTagonist, a kind of Hh signaling inhibitor) in anticancer therapy. However, it is still lacking the
systematic scanning for the common mechanism of various cancer cells in respond to Hh-inhibition.Methodology/Principal Findings: Gene expression profiling of Hh inhibited HT-29 and MKN45 cells was determined by Illumina® Sentrix® BeadChip arrays. From the 17,329 expressed genes across genome, we identified 668 and 269 differentially expressed genes (DEGs, p < 0.01) in comparison pairs of HT-29 and MKN45, respectively. Interesting, large number of common DEGs (77genes) were seen in both two comparison pairs, which was clearly more than the predicted number (10,p < 10−4). Further interpretation of gene ontology was based on over-representation analysis. Two nested categories of GO biological processes (“cell death”, and “response to stimulus”) were detected as candidates with the enrichments of DEGs in both two cell lines. In addition, cDNA microarray profiling of extra cancer cells (ES2 and H4) verified the change of expression in six common DEGs
related to “cell death” (CDKN1A, DDIT3, IER3, IL8, MFGE8, and PPP1R15A) following GANT61 treatment. Conclusions/Significance: Our research indicates that inhibition of Hh has considerable effect on genes from pathway of “cell death” in various carcinoma cells. And, the aberrant of “response to DNA damage” related genes (such as DDIT3) may play a critical role in GANT61-induced cell death. In summary, this dataset provide insight into the molecular mechanisms of GANT61-induced-antitumor activity, and the list of novel GLI-targets in cancer cells.
Key words: Hedgehog signaling pathways; GANT61; Differentially Expressed Gene; anticancer herapy