结直肠癌转移相关基因PCR芯片的建立及初步应用

摘要

Objective To construct a PCR chip with a gene panel for predicting and diagnosing metastatic colorectal cancer. Methods The PCR chip was constructed by integrating 29 genes related to colorectal cancer metastasis identified by gene chip analysis and 3 housekeeping genes into a gene panel. The PCR chip was used for detecting the mRNA expressions of the integrated genes in colorectal cell lines, cancerous specimen and adjacent normal mucosa. The primers for amplification were refined and optimized by several rounds of preliminary reactions. Results The PCR chip containing the 29 candidate genes and 3 housekeeping genes was successfully constructed, which showed specific amplifications of the genes. The results of the PCR chip for detecting the mRNA of the 29 genes related to colorectal cancer metastasis showed a concordance rate of 86% (25 out of 29) with the gene chip data. Application of the PCR chip in the examination of the clinical specimens identified 15 differentially expressed genes between metastatic colorectal cancer and colorectal cancer without metastasis. Conclusion The constructed PCR chip is reliable in the prediction of metastasis of colorectal cancer, and provides a molecular means for evaluating the prognosis of colorectal cancer metastasis.%目的 建立同时检测表达多个结直肠癌转移相关基因的PCR芯片,用于诊断结直肠癌转移和预测转移.方法 通过严格设计基因引物,探索各基因同时达到特异性扩增的PCR反廊条件;应用基因芯片筛选获得的29个结直肠癌转移相关基因,结合3个看家基因,建立同时检测32个基因的PCR芯片;应用该芯片检测不同转移潜能的结直肠癌细胞和临床直肠癌组织、癌旁正常粘膜.结果 成功构建同时检测32个基因的PCR芯片,各基因均获得特异性扩增;29个转移相关基因PCR芯片检测结果与基因芯片检测结果的吻合率为86%;PCR芯片检测伴有转移的结直肠原发癌与不伴有转移的结直肠癌组织,结果显示14个基因表达一致,15个基因表达有差异.结论 结肠癌转移PCR芯片结果可靠,可用于预测结直肠癌转移的研究.