The Study On Interference Therapy In Signal Transduction Of Laryngeal Squamous Cell Carcinoma

The complicated access of signal transduction control all sorts of cell fuctions. Accurate regulation of signal transduction is a hypothesis of life, dysfunction of signal transduction wii lead to pathological process.EGFR and P16 gene take an important part of signal transduction of laryngeal carcinoma. simultaneously,EGFR higher expression and homazygous absence or mutation of P16 present in laryngeal carcinoma.Thus,The study of interference therapy in signal transduction to arm directly at Ad-AS/Ad-pl6 gene stared.To search a original processs for individual gene therapy is our desire.We constructed EGFR antisence cDNA which have complementation to combine domain of EGFR,and constructed pl6β(total length) gene with the AdEasy?vector system in order to study the molecule mechanism of signal transduction of laryngeal carcinoma and aspection,apoptosis,common morphous, ultrastrucure morphous of laryngeal cell.In this process we did EGFR sence cDNA too.At the same time,we raise nude mouse animal model of human laryngeal carcinoma cell-Hep-2. Each group of transplanted tumor respectively directly inject tissu with recombinant adnovirus carried pl6β,AS,S,AS+P16 gene, empty vector and PBS after randomization.Then gross tumor volume change and protein expression or inhibition of target gene were detected in the course of therapy.The data were given statistical treatment with SPSS 11.05 software. The results as follow:1. p16β(total length cDNA), EGFR antisence and sence cDNA (1032bp)were constrcted successfully in our study .moreover, we obtained high titer recombinant adenovirus.2. Recombinant Adenovirus carried target genes infect culture cell Hep-2 efficiently and the protein expression or inhibition of target gene in thesecells were detected.3. After infection of recombinant Adenovirus , culture cell Hep-2 were treated with MTT,FCM,IH,Western Blot etc.The results show target genes can supress actively or kill culture cell Hep-2 except Ad-S.more result show the effect of target genes will enhance if they are combined with each other.4. Infected culture cell Hep-2 were blocked in cyccle G0/G1 stage more than 70% and apoptosis rate of culture cell Hep-2 raiseed, group of Ad-AS+Ad-pl6p is excellent.5. We raise nude mouse animal model in high performance and inject directly target genes in transplanted tumor in order to compare action with target genes.The transplanted tumor were inhibited in some level.The target genes had expressed in transplanted tumor tissu.6. Cell Nucleus in transplanted tumor tissu generated a serise of pathological change and apoptosis underTEM(transmission electron microscope).Conclusion:When the target genes recombinant adenovirus carried were transferred into culture cell Hep-2 and their protein will express or be inhibited effectively.Thus, molecule mechanism of signal transduction of laryngeal carcinoma were interfered successfully.most of tumor cells were blocked GO/Glstage.If we combine Ad-EGFR antisence withAd- P16 the supression of transplanted tumor or culture cell will be all the more.