NGX6对结肠癌细胞系HT-29基因表达谱的影响

Effect of NGX6 on Gene Expression Profile of Colon Carcinoma Cell Line HT-29

背景与目的:结肠癌是最常见的恶性肿瘤之一,其发病机制仍未完全明了。本研究在前期工作的基础上探讨候选抑瘤基因NGX6对结肠癌细胞系HT-29基因表达谱的影响。方法:通过脂质体介导的基因转染构建稳定表达NGX6的结肠癌细胞系pcDNA3.1(+)/NGX6/HT-29,以NGX6转染组为实验组,空白质粒转染组为对照组,利用高通量的表达谱芯片BiostarH-80sx1筛选差异表达基因,RT-PCR验证部分基因芯片的实验结果。结果:NGX6基因转染结肠癌细胞系HT-29后引起该细胞系基因表达谱广泛地改变,其中表达差异3倍以上的基因共377个,这些差异表达基因的功能涉及多个方面,包括细胞信号转导﹑细胞周期调控﹑细胞凋亡﹑细胞骨架和运动﹑基因转录和翻译、DNA损伤修复﹑蛋白质合成和代谢等。其中包括一些非常重要的信号转导通路上的分子改变,如Wnt/β-catenin信号通路上的DKK1、ILK、MMP1、COL11A1,ILK信号通路上的ILK,Eph-Ephrins信号通路上的EpHB2,RhoA信号通路上的ROCK2,RanGTPase信号通路上的RANBP1,以及RB/RBBP1信号通路上的RBBP1等。结论:NGX6转染引起了一些非常重要的信号转导通路上的分子改变,可能通过参与调节这些信号转导通路而发挥其抗肿瘤增殖和转移的作用。

BACKGROUND ＆ OBJECTIVE： Colon cancer is one of the most common malignant tumors, but the mechanism involved in its pathogenesis is not fully understood. Under the foundation of early study, we intended to explore the effects of NGX6 on the gene expression profile of colon carcinoma cell line HT-29. METHODS： pcDNA3.1-NGX6-transfected HT-29 cells were used as the test, and empty vector-transfected HT-29 cells were used as the control. Differentially expressed genes were screened with high-throughout BiostarH-80sxl DNA microarray; part of the results of DNA microarray was confirmed by reverse transcription-polymerase chain reaction （RT-PCR）. RESULTS： NGX6 transfection induced wide changes of the gene expression profile of HT-29 cells. A total of 377 genes were up-regulated or down-regulated by more than 3 folds, which were involved in cell signal transduction, cell cycle controlling, cell apoptosis, cytokinetics, gene transcription and translation, DNA damage and repairing, protein synthesis, metabolism, and so on. Some of them belonged to some important signal pathways related with cell proliferation and metastasis, such as DDK1, ILK, MMP-1 and COL11A1 in Wnt/β-catenin signal pathway, ILK in ILK signal pathway, EpHB2 in Eph-Ephrins signal pathway, ROCK2 in RhoA signal pathway, RANBP1 in RanGTPase signal pathway, and RBBP1 in RB/RBBP1 signal pathway. CONCLUSION： NGX6 transfection leads to molecular changes of some important signal pathways, and may suppress cell proliferation and metastasis of tumor by regulating these signal pathways.