摘要
目的 用基因芯片技术研究人胃腺癌 5 氟尿嘧啶 (5 FU)耐药细胞株SGC7901 /R及其亲本细胞株SGC7901JAK/STAT信号传导通路相关基因表达谱的差异。方法 分别抽提两种细胞mRNA,经逆转录反应,用生物素标记的 16-dUTP将两种细胞的mRNA分别标记成两种cDNA探针,并与载有一组靶基因的人JAK/STAT信号通路芯片进行杂交,通过软件对扫描图像进行数字化处理和分析,筛选 2种细胞在该信号通路中有表达差异的基因。通过RT PCR法分别对耐药细胞及其亲本细胞中的Stat3、核因子(NF)-κB1和bcl-xmRNA的表达水平进行检测。结果 通过两种细胞株JAK/STAT信号通路的基因谱平行比较,筛选出人胃腺癌 5-FU耐药细胞株及其亲本细胞株中有表达差异 2倍以上的基因共 70个,其中表达上调>2倍的 40个,表达下调>2倍的 30个。RT-PCR的验证结果与芯片的筛查结果基本相符。结论 人胃腺癌 5- FU耐药细胞株中JAK/STAT信号通路基因表达谱的变化研究是对胃癌多药耐药分子机制研究的有力补充。
Objective To investigate the difference of gene expression in two human gastric adenocarcinoma cell lines, 5-fluorouracil resistant cell line SGC7901/R and its parental cell line SGC7901, and to screen related genes of JAK/STAT signaling pathway by gene chip. Methods The mRNAs of two cell lines were extracted and purified. The two cDNA probes were made from these two mRNAs which were labelled by (biotin-)16-dUTP, hybridized with human JAK/STAT signaling pathway gene array and scanned for intensity, respectively. The acquired images were analyzed by software. Different expression genes were then screened out. The mRNA expressions of Stat3, NF-κB1 and bcl-x were analyzed by semi-quantitative RT-PCR, (respectively.) Results A parallel comparison between the gene profiles of JAK/STAT signalling pathway in two cell lines showed that a total of 70 genes were screened out whose expressive level was more than 2 folds in 5-flurouracil resistant cell line SGC7901/R and its parental cell line SGC7901. Among these genes, 40 were upregulated and the other 30 were down-regulated. The results of RT-PCR were consistent with those of microarray scanning. Conclusions The knowledge of gene expression profile of JAK/STAT signaling pathway, which was changed in 5-fluorouracil resistant cell line of human gastric adenocarcinoma, has proven to be useful for illustrating the multi-drug resistance mechanisms of gastric cancer.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2005年第1期32-36,共5页
Chinese Journal of Digestion
基金
上海市科委基金资助项目(024119114)
