摘要
目的观察肝素结合性表皮生长因子样生长因子(HB-EGF)激活大鼠肝星状细胞(HSC—T6)过程中微小RNA(miRNA,miR)-221、miR-222表达的变化,探讨miR-221/222在肝星形细胞(HSC)活化过程中的作用。方法培养、接种HSC—T6,分别以HB—EGF和细胞外信号调节激酶(ERK)特异性抑制剂PD98059+HB—EGF共处理HSC—T624、48h,采用实时定量反转录聚合酶链反应(RT—qPCR)法检测miR-221/222表达水平。结果对照组、HB—EGF组及共处理组(48h)miR-221表达水平分别为0.90±0.14、1.08±0.09和0.71±0.14,miR-222表达水平分别为0.82±0.10、1.04±0.09和0.68±0.16。说明HB—EGF可促进miR-221、miR-222的表达,ERK抑制剂可使miR-221、miR-222表达下降(P〈0.01)。结论HB—EGF通过激活HSC—T6的Ras/ERK信号通路,继而促进miR-221、miR-222的表达,参与HSC活化的调控。
Objective To study the effect of heparin - binding epidermal growth factor (HB -EGF) on the expression of microRNA (miRNA, miR) -221/222 for exploring the regulatory roles of miR -221/222 in the activation of hepatic stellate cells (HSC). Methods The rat HSC -T6 cells were treated with HB - EGF and PD98059 HB - EGF for 24 or 48 h. The expression of miR - 221 and miR -222 mRNA was detected by real- time quantitative reverse transcriptase -polymerase chain reaction ( RT - qPCR). Results RT - qPCR showed the level of miR - 221 in control group, HB - EGF group and co -cultured group was 0. 90 ±0. 14, 1.08 ±0. 09 and 0. 71±±0. 14 respectively, and that of miR -222 was O. 82 ±0. 10, 1.04 ± 0. 09 and 0. 68 ± 0. 16 respectively. The expression levels of miR - 221 and miR - 222 were significantly increased in HSC - T6 cells treated with HB - EGF, and those were decreased in HSC - T6 cells co - cultured with PD98059 and HB - EGF, as compared with control group ( P 〈 0. 01 ). Conclusion HB - EGF participates in the regulation of HSC activation by activating Ras/Extracel- lular sigual regulated kinase (ERK) signal transduction pathway in HSC -T6 cells and subsquently accel- erating the expression of miR- 221/222.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第12期3032-3034,共3页
Chinese Journal of Experimental Surgery
基金
福建省临床医学重点专科资助项目(闽卫科教[2012]149号)