The Expression And Correlation Research Of MicroRNA-10a-5p And PIK3CA In Middle Ear Cholesteatoma

Objective: To study the expression differences,distribution and the correlation of micro-RNA-10a-5p(miR-10a-5p)and it’s target gene phosphatidy-linositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA)protein in cholesteatoma tissues.This study aims to provide new research ideas and directions for the non-surgical treatment of middle ear cholesteatoma by exploring the role of miR-10a-5p and PIK3CA in the molecular biological mechanism of the occurrence and development of middle ear cholesteatoma.Methods: 1.A total of 27 cases of cholesteatoma and normal skin tissues behind the ear were collected.The experimental group was cholesteatoma tissue of the middle ear,and the control group was normal skin tissue behind the ear.2.PIK3CA was predicted to be the downstream target gene of miR-10a-5p by Target Scan,miRBase and other bioinformatics software.3.Quantitative Real-time PCR(qRT-PCR)was used to detect the mRNA expression of miR-10a-5p and PIK3CA in the cholesteatoma and the normal skin tissues behind the ear.4.Immunohistochemistry and Western Blot were used to detect the expression of PIK3CA in middle ear cholesteatoma and normal skin tissue behind ear.5.The correlation between miR-10a-5p and PIK3CA mRNA expression in cholesteatoma tissues was analyzed by statistical methods to further explore whether miR-10a-5p participates in the growth and proliferation of middle ear cholesteatoma epithelial tissues by regulating the function of PIK3CA.Results: 1.QRT-PCR results indicated that the expression of miR-10a-5p could be detected in cholesteatoma and normal skin tissues behind the ear,and the expression level of miR-10a-5p in cholesteatoma(0.99±0.03)was significantly lower than that in normal skin tissues(1.61±0.32),with statistical significance(P < 0.05).2.QRT-PCR results showed that PIK3CA mRNA expression could be detected in cholesteatoma group and normal skin tissues behind the ear,and the expression level of PIK3CA mRNA in cholesteatoma(2.06±0.23)was significantly higher than that in normal skin tissues(1.02±0.05),with statistical significance(P < 0.05).3.Immunohistochemical results indicated that the expression of PIK3CA in cholesteatoma tissues was mainly localized in the cytoplasm and cell membrane,with positive expression from basal layer to basal layer,and the staining degree was mostly moderate to strong positive.PIK3CA was mainly localized in the nucleus and cell membrane in the normal skin tissues behind the ear,showing staining in the basal layer and a few basal superlayer cells,and the degree of staining was mostly weak positive.4.Western Blot results showed that the expression level of PIK3CA in cholesteatoma tissues was significantly higher than that in the control group of normal skin tissues,with statistical significance(P < 0.05).5.Pearson correlation analysis showed that miR-10a-5p was significantly negatively correlated with PIK3CA in cholesteatoma tissues of middle ear(r=-0.926,P <0.001).Conclusions: 1.The lower expression of miR-10a-5p in middle ear cholesteatoma tissues may inhibit the growth and proliferation of cholesteatoma.2.The higher expression of PIK3CA in middle ear cholesteatoma tissues may promote the growth and proliferation of cholesteatoma.3.MiR-10a-5p was negatively correlated with PIK3CA expression in middle ear cholesteatoma tissues,suggesting that down-regulated miR-10a-5p may affect the proliferation and differentiation of cholesteatoma keratinocytes by negatively regulating the function of its target gene PIK3CA.