用原位RT-PCR技术检测大肠癌组织中相关新基因SNC66的表达

Detecting the expression of SNC66 gene in colorectal cancer tissue using In situ reverse transcription and polymerase chain reaction

目的 :探讨在肿瘤组织石蜡切片标本中原位检测内源性基因表达产物的灵敏方法,研究大肠癌相关新基因SNC66在大肠癌组织中的表达情况以及与大肠癌的相关性。方法 :经原位逆转录后进行原位PCR扩增,在扩增过程中掺入地高辛标记的核苷酸(Dig-UTP)并加以检测。比较SNC66在大肠粘膜和大肠癌组织中的表达情况。结果 :25个循环时,37例大肠癌组织标本有10例阴性不表达,27例强阳性表达。10个循环时,则14例呈阴性,18例弱阳性,5例强阳性。配对的37例大肠粘膜标本则都呈强阳性表达。结论 :原位RT -PCR是一种检测内源性基因低拷贝转录产物mRNA的较灵敏的方法。SNC66基因在大肠癌组织中存在明显的表达降低或缺陷,可作为侯选的抑癌基因加以进一步研究。

Objective: To study the a sensitive non-isotope method for in situ detection on the expression of endogenous genes in paraffin-embeded section of cancer tissue, study the expression of SNC66 genes in colorectal cancer(CRC) and investigate the relationship between the expression of SNC66 gene and the genesis of the tumor. Methods: Paired tumor and normal tissues from the CRC patients were tested by in situ RT-PCR in which the SNC66 fragments were labeled with digoxingen and detected with NBT/BCIP. Results: Ten are negative and 27 are strong positive in 37 cases when the PCR cycles 25 times, and 14 are negative and 18 are weak positive,and 5 are strong positive when the PCR cycles 10 times. Conclusion: The method of in situ RT-PCR for the detection of low copies of endogenous is sensitive and useful, There is a significant expression reduction of SNC66 in CRC tissues.SNC66 should be further investigated as a candidate suppressor gene.