Abstract

 
 
 
 Purpose: To determine the effect of MiR-21 on the proliferation of glioma cells in children with cholesteatoma, and the pathway involved.
 Methods: Cholesteatoma tissues from children with cholesteatoma in Otology Department of our hospital were isolated, extracted and cultured in serum-free medium of keratinocytes. The experiment comprised three groups: negative control group, blank control group and miR-21 inhibition group. Cell cycle and cell proliferation were analyzed. mRNA and protein expressions of phosphatase tension homologue (PTEN) and programmed cell death factor-4 (PDCD4) were determined.
 Results: The proliferation of CK cells in miR-21 group was significantly lower than that in negative control and blank control groups (p < 0.05). The proportion of CK cells at G0/G1 phase in miR-21 inhibition group was significantly higher than those in negative control and blank control groups (p < 0.05). The protein and mRNA expression levels of PTEN and PDCD4 in CK in miR-21 group were significantly higher than those in negative control and blank control groups (p < 0.05).
 Conclusion: Inhibition of miR-21 slows down cell proliferation to some extent, and induces apoptosis of cholesteatoma cells by inducing cell cycle arrest at G0/G1 phase via a mechanism linked to the negative regulation of expressions of PTEN and PDCD4. Thus, MiR-21 can be used as potential target for the drug of treatment of children with cholesteatoma.
 
 
 
Highlights
Cholesteatoma of the middle ear, a relatively common and frequently occurring disease, is an abnormal accumulation of keratin-producing squamous epithelium in the middle tympanum, superior tympanum, and mastoid process or rocky apex [1]
There was no significant difference in % cell proliferation between the negative control group and blank control group (p > 0.05)
The proportion of CK cells in G0/G1 phase in the miR-21 inhibition group was significantly higher than the corresponding values in the negative control and blank control groups, while the proportion of cells in S phase was significantly lower than those in the negative control and the blank control groups. These results suggest that miR-21 inhibition induces apoptosis of cholesteatoma cells by inducing arrest of cholesteatoma cell cycle at the G0/G1 phase
Summary
Cholesteatoma of the middle ear, a relatively common and frequently occurring disease, is an abnormal accumulation of keratin-producing squamous epithelium in the middle tympanum, superior tympanum, and mastoid process or rocky apex [1]. Cholesteatoma in children has a rapid growth rate and a wide range of lesions, and it spreads with significantly higher postoperative recurrence than cholesteatoma in adults [4]. It has been found that miRNA-21 is significantly overexpressed in cholesteatoma tissue, and may be involved in the etiology of cholesteatoma cells [7]. Cholesteatoma cells were used to study the effect of miR-21 on the proliferation of glial cells in children cholesteatoma, as well as the related mechanisms. Biological Experimental Equipment Co., while high speed table top centrifuge was obtained from Beijing Ganming Gene Technology Co. Ltd. In the blank control group, 200 μL of PBS was added to each well. 100 μL RNASeA was added in 37 °C water bath for 30 min, followed by staining with 200 μL PI in the dark at 4 °C for 30 min. The red fluorescence was recorded at the excitation wavelength of 488 nm
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
