Chen Yao, Zhang Yi-Zeng, Zhou Zong-Guang, Wang Gang, Yi Zeng-Ni
Department of Anatomy, Basic and Legal Medical Institute, West China Medical Center, Sichuan University, Chengdu 610041, Sichuan Province, China.
World J Gastroenterol. 2006 Feb 21;12(7):1025-32. doi: 10.3748/wjg.v12.i7.1025.
To investigate the differently expressed genes in human colorectal adenocarcinoma.
The integrated approach for gene expression profiling that couples suppression subtractive hybridization, high-throughput cDNA array, sequencing, bioinformatics analysis, and reverse transcriptase real-time quantitative polymerase chain reaction (PCR) was carried out. A set of cDNA clones including 1260 SSH inserts amplified by PCR was arrayed using robotic printing. The cDNA arrays were hybridized with fluorescent-labeled probes prepared from RNA of human colorectal adenocarcinoma (HCRAC) and normal colorectal tissues.
A total of 86 genes were identified, 16 unknown genes and 70 known genes. The transcription factor Somult9 influencing cell differentiation was downregulated. At the same time, Heat shock protein 10 KDis downregulated and Calmoulin is up-regulated.
Downregulation of heat shock protein 10 KD lost its inhibition of Ras, and then attenuated the Ras GTPase signaling pathway, increased cell proliferation and inhibited cell apoptosis. Down-regulated transcription factor Somult9 influences cell differentiation and cell-specific gene expression. Down-regulated Somult9 also decreases its binding to calmodulin, accumulates calmodulin as receptor-activated kinase and phosphorylase kinase due to the activation of PhK.
研究人类结肠腺癌中差异表达的基因。
采用抑制性消减杂交、高通量cDNA阵列、测序、生物信息学分析和逆转录实时定量聚合酶链反应(PCR)相结合的基因表达谱综合方法。使用机器人打印技术将一组包括通过PCR扩增的1260个SSH插入片段的cDNA克隆进行阵列排列。cDNA阵列与用人结肠腺癌(HCRAC)和正常结肠组织的RNA制备的荧光标记探针杂交。
共鉴定出86个基因,其中16个未知基因和70个已知基因。影响细胞分化的转录因子Somult9下调。同时,热休克蛋白10KD下调,钙调蛋白上调。
热休克蛋白10KD的下调失去了对Ras的抑制作用,进而减弱了Ras GTPase信号通路,增加了细胞增殖并抑制了细胞凋亡。转录因子Somult9的下调影响细胞分化和细胞特异性基因表达。下调的Somult9还减少了其与钙调蛋白的结合,由于磷酸化酶激酶(PhK)的激活,导致钙调蛋白作为受体激活激酶和磷酸化酶激酶积累。