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目的研究抗癌活性肽对人胆囊癌细胞系GBC-SD细胞作用前后的基因表达谱,筛选与抗癌活性肽作用相关的基因表达改变。方法将1152条人类全长基因的PCR产物按微矩阵排列点样于特殊处理的玻片上制备成表达谱芯片;按一步法抽提抗癌活性肽作用前后体外培养的胆囊癌细胞的RNA并纯化mRNA,通过逆转录用两种荧光Cy3-dUTP和Cy5-dUTP标记对照和实验组胆囊癌细胞cDNA,制成cDNA探针并与表达谱芯片杂交,用ScanArray4000扫描仪扫描芯片上两种荧光信号,计算机分析判定差异表达的基因。结果在1152条基因中筛选出有差异表达的基因245条,其中上调的121条,下调的124条。结论利用本基因表达谱芯片可同时研究数以千计的基因表达水平,从而在基因水平上探讨抗癌活性肽对胆囊癌GBC-SD细胞作用的机制。
Objective To study the gene expression profile of anticancer active peptides on human gallbladder carcinoma cell line GBC-SD cells before and after treatment, and to screen the gene expression changes related to anticancer active peptides. Methods PCR products of 1152 human full-length genes were micro-arrayed on special treated glass slides to prepare expression microarray. The RNA of gallbladder carcinoma cells cultured in vitro was extracted and purified by one-step method mRNA, reverse transcription using two fluorescent Cy3-dUTP and Cy5-dUTP labeled control and experimental group of gallbladder cancer cDNA, made of cDNA probes and hybridization with the expression profiling chip ScanArray4000 scanner with two kinds of fluorescence signals on the chip, the computer Analysis of differentially expressed genes. Results Among the 1152 genes, 245 differentially expressed genes were screened out, of which 121 were up-regulated and 124 were down-regulated. Conclusion The gene expression profiling chip can simultaneously study thousands of gene expression levels to explore the mechanism of action of anticancer active peptides on GBC-SD cells at the gene level.