The Screening Of Cytokine Biomarkers And Its Significance In Recurrent Nasal Polyps

Chapter 1The study of the biomarkers in JAK-STAT, NF-κB and MAPK signaling pathway of the recurrent nasal polyps tissues with functional gene chips techniqueObjective:To detect the differentially expressed genes in the signaling pathway of JAK-STAT, NF-κB and MAPK in the recurrent nasal polpys through functional gene chips technique and evaluate the significance of these detected genes in the pathogenesis and recurrence of nasal polyps.Methods:The cDNAs were synthesized from total RNA which were extracted from 4 groups (10 cases respectively) including nasal polyps, recurrent nasal polyps, chronic sinusitis and normal nasal mucosa subjects. The cDNA were then taken into standard 96-well PCR equipment for real time PCR. The fluorescence signals of gene chip were obtained through a scanner. The expression difference of compared genes between experimental and normal groups in singaling pathway of JAK-STAT,MAPK and NF-κB was calculated by 2-ΔΔCt. These experiment processes were repeated strictly accorded to the experiment design to assure the stability and reliability of the final data.Results:There were 14 differentially expressed genes which were up-regulated up to more than 2-fold compared with the normal group, including 9 genes in JAK-STAT signaling pathway,2 genes in MAPK signaling pathway and 3 genes in NF-κB signaling pathway. These differentially expressed genes were singal transduction and transcription activation factor, transcription factor, cytokine transduction inhibition factor, epidermal growth associated factor, cell cycle regulation factor and complement factor, etc. Among the 14 candidates, there are seven genes whose upregulation may specially reflects the recurrent status of the nasal polyps, four genes for the status of nasal polpys and one for the inflammation status of nasal mucosa. Conclusions:It is feasible to screening the differentially expressed genes among the nasal polyps, recurrent nasal polyp, chronic sinusitis and normal nasal mucosa by means of functional classification gene chip technique. It will provide a new method in development of the objective experimental indicator for the classification and prognosis evaluation of nasal polyps.Chapter 2The expression and significance of Cyclin D1 and Oncostatin M in the recurrent nasal polypsObjective:To detect the expressions of Cyclin D1 and Oncostatin M proteins in recurrent nasal polyps, nasal polyps, chronic sinusitis and normal nasal mucosa so as to study their roles in the pathogenesis of the recurrent nasal polyps.Methods:The protein expression of Cyclin D1 and Oncostatin M were detected by Western blotting in recurrent nasal polyps, nasal polyps, chronic sinusitis and normal nasal mucosa cases (6 cases for each group). The expression and localization of Cyclin D1 were immunohistochemi-cally observed in the above four groups.Results:(1) Cyclin D1 protein was expressed in all of the specimen of the four groups, and predominantly stained in the nuclei of both epithelial cells of nasal mucosa and the underlying gland cells. The positive staining was yellow or claybank. By western blotting the expressions of Cyclin D1 protein and OSM protein in the recurrent nasal polyps, nasal polyps and chronic sinusitis were higher than that in normal middle turbinate mucosa. (2) The protein expressions of OSM and Cyclin D1 in the recurrent nasal polyps were significantly higher than that in other groups (p<0.05).Conclusions:The expressions of Cyclin D1 and OSM were up-regulated in the tissue of recurrent nasal polyps, which indicate that they may play the important roles in the development of recurrent nasal polyps, and may be utilized as the candidate biomarkers for detecting the susceptibility of recurrent nasal polyps.