肝素结合性表皮生长因子样生长因子对大鼠肝星状细胞-T6表达微小RNA-221、微小RNA-222的影响

Effect of heparin - binding epidermal growth factor on the expression of microRNA - 221/222 in HSC- T6

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.