Cloning, Screening, Location And Function Study Of Oral Lichen Planus Related Genes

Oral lichen planus (OLP) is one of common chronic oral mucosal diseases, and its pathogenesis has not been clear until now,moreover, the effect of its clinical treatment has not been good,so the heath and life of patients has been affected badly by this disease. OLP has been recognised as precancerous change by world hygeine organization because of its tendency to be cancer. There has been no systematic studies on it, however, some following studies showed that the proliferation of cells with immunoactivities, the expression change of cellular growth factor, apoptosis and the specific expression of proliferation related genes played an important role on the occurrence of OLP. The change and interaction of multigene tend to cause some disease, therefore, the investigation into gene expression of pathological tissue will contribute to the elucidation of pathogenesis. A subtractive hybridization method is adopted in this article,and mRNA is isolated directly from pathological tissue of patients with OLP and their normal tissue, and different expressing genes related OLP are cloned and screened, then the tissue expression of some genes is investigated. This work is composed of two parts. 1 Clone and screening of OLP related genesThe purpose of this study is to construct a subtractive cDNA library ofgenes expressed differently between unormal tissue of OLP and normal tissue of one patient. The mRNA was extracted respectively from the unormal and normal tissue, and the cDNA library with a capacity of two hundred was constructed by an improved technique based on PCR subtractive hybridization. One hundreds clones were randomly selected and identified with restriction enzymes,64 clones was found including an insert fragment of 500 bp. Reverse dot blot hybridization was used to identify and get rid of false positive clones,then 16 clones was selected randomly to be sequenced and analysed,and the results showed that these genes cound be divided into three types,such as cellular growth related genes,bone development and metabolism related genes,and inflammation immunity signal transduction related genes. TRAF-6 is one of the cloned gene,which is a regulation gene related cellular growth and inflammation immunity signal transduction. TRAF-6 is not only regulate cellular apoptosis,also induce some genes to express protein contribute to cellular subsistence. As a junction protein, TRAF-6 can interact with different kinds of cellular membrane receptor,and play an important role on the regulation of cellular apoptosis.TRAF-6 is involved in the regulation of cellular proliferation and apoptosis by signal transduction. We investigated the expression of ETS-1,TRAF-6 and TNF-cc in the oral mucosa with OLP and normal oral mucosa,and the result showed that both TRAF-6 and TNF-aexpressed strongly in mucosa with OLP, moreover,their expression was direct proportion. The specific tissue expression of ETS-1,TRAF-6 inferred that it might be OLP related gene.