| Objective:The expression of proliferating cell nuclear antigen(PCNA),keratinocyte growth factor(KGF / FGF7)and keratinocyte growth factor receptor(KGFR / FGFR2)in 29 cases of cholesteatoma and 13 cases of chronic suppurative otitis media and 10 cases of normal external auditory canal skin were detected.To investigate the expression of three kinds of molecules in acquired cholesteatoma and chronic suppurative otitis media and their relationship and differences.The clinical feature and the symptoms of cholesteatoma and chronic suppurative otitis media were classified and analyzed.The possible mechanism of ossicular chain damage,hearing loss,bone erosion are were discussed which may shedding lights on non-invasive biological targeting therapy for clinic.Methods:The 42 patients with otitis media from Department of Otorhinolaryngology of the first affiliated Hospital of Southwest Medical University from July 2017 to December 2017 were collected,After operation,42 cases were confirmed by pathological examination.There were 13 cases of inflammatory granulation and 29 cases of cholesteatoma,another 10 cases of normal skin trimmed in external auditory canal cavum conchaeplasty were taken as control.Clinical matierals of 42 patients were also reviewed,including medical records,ear endoscopy,HRCT and MRI in some patients.The experiment was divided into three groups: cholesteatoma group(group 1),otitis media group(group 2)and normal skin group(group 3).One of them was stained with routine hematoxylin-eosin(HE)or Giemsa staining for morphological observation and confirmation.The expression and localization of the three molecules were detected by immunohistochemical SP method(streptavidin labeled by peroxidase).Immunohistochemical staining section was used to collect 8 unoverlapped visual field images of positive expression region under OLYMPUS microscope.The mean optical density of each image was calculated by image pro plus 6.0 software.The mean value of MOD of 8 images was taken as the expression amount of three kinds of molecules in each slice.According to the expression of KGF/KGFR/PCNA in three groups,combined with the clinicopathological features of cholesteatoma and chronic suppurative otitis media,all the data were analyzed by SPSS17.0.Results: the MOD value in Statistics(mean ±SD):(1)the KGF was mainly located in the cytoplasm,membrane,also found in nucleus in cholesteatoma epithelium,inflammatory granulation tissue and normal skin of the average MOD were(0.222±0.048),(0.118±0.044),(0.106±0.017),the highest expression of cholesteatoma,3 group samples were compared using one-way ANOVA,F = 40.496,P =0.000,it’s showed that the total expression of KGF was different in the 3 groups,and was statistically significant(P < 0.05).Further comparison of LSD analysis showed that the expression of cholesteatoma and inflammatory granulation,.In cholesteatoma group,there was significant difference between normal skin and inflammatory granulation(P < 0.05),but there was no difference between inflammatory granulation and normal skin(P > 0.05).the results of SNK showed that the expression of inflammatory granulation was in the same column as normal skin.The cholesteatoma group and the other two groups were not in the same column,so there were differences.The results were consistent with LSD method.(2)The mean MOD of KGFR was(0.242±0.058),(0.109±0.010),(0.106±0.009),P = 0.000,which indicating that there was significant difference in the expression of KGFR among the three groups(P < 0.05).The results of LSD showed that there was significant difference between the cholesteatoma group and the normal skin group in expression of KGFR(P < 0.05),but there was no difference between the inflammatory granulation group and the skin group.The results of 0.05).SNK were consistent with LSD method.(3)PCNA is mainly located in the nucleus.The average MOD of the three groups were(0.372±0.067),(0.266±0.032),(0.237±0.020)respectively.The highest expression of PCNA was found in cholesteatoma group(F = 31.997,P=0.000).It can be concluded that there is a statistical difference in the expression of PCNA between and within the three groups(p < 0.05).The results of LSD showed that there was statistical difference between cholesteatoma and inflammatory granulation or normal skin group,but there was no significant difference between inflammatory granulation and skin group.The results of SNK method showed that cholesteatoma group higher than inflammatory granulation and normal skin.The skin does not differ from the same subset,The result of detection was the same as that of LSD method.(4)According to the clinical matierals of cholesteatoma and chronic suppurative otitis media,the expression of three kinds of molecules were classified and analyzed: the three molecules were strongly expressed in cholesteatoma group,followed by chronic suppurative otitis media granulation group;By pearson correlation analysis,it was found that there was no significant correlation between the three molecular expressions about age,sex,ear side,duration of disease,clinical symptoms(P > 0.05),but in cholesteatoma and chronic suppurative otitis media with severe hearing loss.Bone destruction of ossicular chain,the expression was higher than patients with tender bone erosin or hearing loss(P < 0.05),which with positively correlation(P < 0.05).(5)The expression of three kinds of molecules in cholesteatoma group was significantly higher than that in inflammatory granulation and normal skin group.There was no significant difference between normal skin and inflammatory granulation group.Scatter plot showed that distribution of the three kinds of molecular expression was approximately linear correlation,and then compared with each other by pearson correlation analysis.It was found that there was a positive correlation between the expression of KGF,KGFR and PCNA in cholesteatoma and granulation.Cholesteatoma group:KGF/KGFR(r=0.759,p=0.000),KGF/ PCNA(r=0.858,p=0.000),KGFR/PCNA(r=0.820,p=0.000).Chronic suppurative otitis media:KGF/KGFR(r=0.863,p=0.000),KGF/PCNA(r=0.813,p=0.001),KGFR/PCNA(r=0.979,p=0.000).conclusion: the expression of KGFR in cholesteatoma epithelium was significantly higher than that in inflammatory granulation tissue and the skin of external auditory canal,suggesting that the three molecules were involved in the process of excessive proliferation of cholesteatoma and played a synergistic role.2.The positive expression of KGF in fibroblasts,chronic inflammatory cells and vascular endothelial cells in cholesteatoma and inflammatory granulation group suggests that KGFR and KGF may also regulate angiogenesis in inflammatory microenvironment.In cholesteatoma and chronic suppurative otitis media in the progress of the disease play a facilitating role.3.The expression of PCNA,KGF,KGFR in cholesteatoma and chronic suppurative otitis media is related to the degree of hearing loss,the degree of ossicular chain damage,the degree and extent of bone destruction.These results suggest that the overexpression of these three molecules may play a synergistic role in the process of invasive growth and bone destruction.4.The deficiency of this study :The overexpression of three molecules of PCNA,KGF,KGFR may be related to the destruction of bone in cholesteatoma and chronic suppurative otitis media,But more basic experimental research and large sample clinical evidence are needed for further examination. |
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