Identification Of The Deleted Genes In Human Esophageal Cancer

Esophageal cancer (EC) is one of the most common cancers in China. During the last 15 years, cytogenetic and molecular studies on human esophageal cancer carried out in our laboratory have revealed consistent chromosomal aberrations and high frequencies of loss of heterozygosity (LOH) and micro satellite instability (MIN) at many loci. All these data are indicating existence of tumor suppressor genes of this human malignancy.Representational difference analysis (RDA) based on genomic subtractive hybridization technique is one of a few new strategies for finding small differences between normal and mutated complex genomes. In contrast to current approaches in positional cloning, RDA has the potential to save a great deal of repetitive work involving analysis of multiple markers and large pedigrees of diseases to require knowledge of functions or positions. Therefore RDA provides an economical and sensitive route to solve many types of genetic problems. However, the procedure is complicated and time-consuming. First, preparation of the tester and the driver involves a series of steps including PCR amplification, digestion ,and recovery of PCR products from low melting agarose. Second, single-stranded endonucleases may digest duplex nucleic acid due to their excess amount and non-specific activities. So some informative materials can be lost.