Effect of microRNA-143 on interleukin-13-induced expression of kallikrein 7 in human epidermal keratinocytes
10.3760/cma.j.issn.0412-4030.2018.04.003
- VernacularTitle:MicroRNA-143对白介素13诱导的人角质形成细胞组织激肽释放酶7表达的影响
- Author:
Yueping ZENG
1
;
Chenyu ZHU
;
Cheng CHEN
;
Qiannan JIA
;
Hongzhong JIN
Author Information
1. 中国医学科学院北京协和医学院北京协和医院皮肤科协和转化医学中心
- Keywords:
Dermatitis,atopic;
Keratinocytes;
Interleukin-13;
Kallikrein 7;
MicroRNA-143
- From:
Chinese Journal of Dermatology
2018;51(4):256-259
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the effect ofmicroRNA-143 (miR-143) on interleukin (IL)-13-induced expression of kallikrein 7 (KLK7) in primary normal human epidermal keratinocytes (NHEKs).Methods Some NHEKs at exponential growth phase were divided into 4 groups to be treated with recombinant human IL-13 at different concentrations of 0,2,10 and 50 μg/L respectively for 24 hours,and some NHEKs were treated with 50 μg/L IL-13 for 0,6,12,24 and 48 hours separately.After the treatment,NHEKs were collected,and total RNA was extracted.Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of KLK7.Some other NHEKs were divided into another 4 groups:NHEK group (blank control group) receiving no treatment,IL-13 group treated with 50 μg/L IL-13,miR-NC group transfected with miRNA mimics negative control followed by the treatment with 50 μg/L IL-13,and miR-143 group transfected with miR-143 mimics followed by the treatment with 50 μg/L IL-13.After 24-hour treatment with IL-13,real-time fluorescence-based quantitative PCR and Western blot analysis were conducted to determine the mRNA and protein expression of KLK7 respectively in the above groups.Results After 24-hour treatment with IL-13 at concentrations of 0,2,10 and 50 μg/L,the mRNA expression of KLK7 in NHEKs was 1.00 ± 0.12,0.89 ± 0.04,1.15 ± 0.09 and 1.70 ± 0.10 respectively,and significantly increased along with the increase of IL-13 concentrations (F =92.48,P ＜ 0.05).After 0-,6-,12-,24-and 48-hour treatment with 50 μg/L IL-13,the mRNA expression of KLK7 in NHEKs was 1.00 ± 0.05,1.05 ± 0.12,1.71 ± 0.20,1.97 ± 0.19 and 2.48 ± 0.13 respectively,and significantly increased over time (F =206.44,P ＜ 0.05).Compared with the miR-NC group,the miR-143 group showed significantly decreased mRNA and protein expression of KLK7 (t =6.76,4.23 respectively,both P ＜ 0.05).Conclusion In NHEKs,IL-13 can up-regulate the expression of KLK7,likely by the regulation of miR-143.
