基因芯片技术筛选结直肠癌腹膜转移相关基因

Application of genome-wide microarray for screening genes related to peritoneal metastasis of colorectal cancer

目的筛选和鉴定结直肠癌腹膜种植转移相关基因。方法收集手术切除3例伴有腹膜种植转移的结直肠腺癌病人原发病灶和正常黏膜的新鲜组织标本,提取总RNA,经逆转录合成cDNA后经体外扩增合成aRNA,Cy3荧光分子标记后和人类全基因组表达谱芯片杂交,采用经验贝叶斯方法(P≤0.05)筛选出差异表达基因,并用RT-PCR实验对部分差异表达基因进行验证。结果满足(P≤0.05)的基因共有105个。其中和正常组织相比,在肿瘤组织表达上调的基因有42个,表达下调的基因有63个。通过生物信息学分析,在表达上调基因中选择3条(S100P;PRDX1;SLPI)基因进RT-PCR行验证,结果与芯片结果完全相符。结论基因芯片技术通过筛选结直肠癌腹膜转移过程中发挥关键作用的基因,为寻找结直肠癌腹膜转移的分子标志物提供依据和参考。

Objective To screen genes related to peritoneal metastasis of colorectal cancer.Methods Specimens of primary cancer and normal mucosa tissues were collected from 3 patients with peritoneal metastasis of colorectal cancer.The total RNA were extracted and inversely transcribed into cDNA to synthesize aRNA using in vitro RNA synthesis.The synthesized aRNA,after labeling with Cy3,were hybridized with the whole human genome oligo microarray.The Empirical Bayes method was used to screen the differentially expressed genes,followed by confirmation of the selected genes by semi-quantitative RT-PCR.Results With a threshold of P≤0.05,a total of 105 differentially expressed genes were identified in primary cancer lesions,including 42 up-regulated and 63 down-regulated genes.Three of the up-regulated genes（S100P,PRDX1 and SLPI） were selected and confirmed by RT-PCR,which yielded results consistent with those from gene microarray.Conclusion Gene microarray technique can provide valuable clues for locating the tumor markers of peritoneal metastasis in colorectal cancer patients.