Differential Expression of miR-133b in Human Hair Follicles Treated with DHT in Vitro and Its Functional Analysis in Human Dermal Papille Cells
- VernacularTitle:双氢睾酮体外培养人毛囊中差异表达的miR-133b对人毛乳头细胞的增殖及诱导能力的影响
- Author:
Wen-jia DENG
1
;
Chang-jian DENG
2
;
Le HAN
1
;
Ben LIU
1
;
Xin TANG
1
;
Miao-jian WAN
1
Author Information
1. Department of Dermatology and Venereology， The Third Affiliated Hospital of Sun Yat-sen University， Guangzhou 510630， China
2. Department of Otorhinolaryngology， the People's Hospital of Longgang District， Shenzhen 518172， China
- Publication Type:Journal Article
- Keywords:
miR-133b;
5α-dihydrotestosterone;
hair follicles;
dermal papille cells
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2021;42(2):202-208
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the effects of different concentrations of 5α-dihydrotestosterone （DHT） on growth and proliferation of human hair follicles （HFs） and their relationship with miR-133b expression， then further explore the role of miR-133b in the proliferation and inducibility of human dermal papille cells. MethodsHFs were isolated by microdissection， then the anagen isolated HFs were cultured and divided into different concentrations of DHT treatment groups （10-8mol/L， 10-7mol/L， 10-6mol/L， 10-5mol/L） and the blank control group. The HF growth and morphology were measured and evaluated. The expression of Ki-67 in hair matrix cells and miR-133b were detected by immunofluorescence assay and qRT-PCR respectively. Lipofectamine 2000 was used to transfect miR-133b mimics and miR-133b NC into human dermal papilla cells. CCK-8 was used to assess the proliferation ability of human dermal papilla cells. qRT-PCR and Western Blot were performed to evaluate respectively the mRNA and protein levels of the markers associated with inductive ability of dermal papilla cells， such as Versican， ALP and β-catenin. ResultsCompared with the control group， no other treatment groups but the DHT 10-5mol/L group showed statistically significant inhibitory effect on HF growth （P<0.05）. The catagen in the DHT 10-5mol/L group appeared earlier than that in the control group and there was no statistically significant difference in HF growth between other treatment groups and the control group. The DHT 10-5mol/L group showed lower percentage of Ki-67-positive cells in hair matrix cells and significantly increased relative expression of miR-133b in HFs， 3.17±0.26 times more than that in the control group （P<0.01）. CCK-8 assay revealed that the OD450 value in the miR-133b mimics group was lower than that in the miR-133b NC group （P<0.05）. qRT-PCR revealed that the mRNA expression levels of Versican， ALP， and β-catenin in the miR-133b mimics group were all lower than those in the miR-133b NC group （P<0.001， P<0.01， P<0.01）. Western Blot revealed that the protein expression levels of Versican， ALP， and β-catenin in the miR-133b mimics group were also lower than those in the miR-133b NC group （P<0.01， P<0.001， P<0.01）. ConclusionsHigh concentrations of DHT may inhibit the growth and proliferation of HFs via regulating the expression of miR-133b， thus affect the proliferation and inducibility of human dermal papilla cells.
