摘要
【目的】探讨mi R-200c在肝星状细胞(HSC)活化、增殖、迁移及促肝纤维化中的作用。【方法】慢病毒转染的方法构建过表达mi R-200c的稳定细胞株(LX2-200c)和空质粒对照组(LX2-nc),RT-PCR检测LX2-200c和LX2-nc细胞上mi R-200c和Ⅰ型胶原蛋白的表达,Western Blot检测LX2-200c和LX2-nc细胞的活化标志分子α-SMA和Vimentin的变化,CCK-8实验检测增殖情况,划痕实验检测迁移情况。【结果】野生型LX2细胞中mi R-200c呈低表达,mi R-200c的过表达促进了HSC上α-SMA和Vimentin表达的增加,并使HSC的增殖和迁移能力增强,另外,ECM重要组成成分Ⅰ型胶原蛋白的表达也显著增加(P=0.0031)。【结论】mi R-200c可以促进HSC的活化、增殖、迁移以及ECM的分泌,进而参与了肝纤维化的过程。
[ Objective ] To investigate the role of miR-200c in activity, proliferation, migration and pro-fibrosis of hepatic stellate ceils (HSC). [Methods] Human hepatic stellate cell lines (LX2) with stably expressing miR-200c (LX2-200c) and empty vector control (LX2-nc) were constructed successfully by lentiviral vector system respectively. The level of miR-200c and collagen type I in LX2-200c and LX2-nc cells were detectedusing real time PCR.The variation of the activation markers (ct-SMA, Vimentin) in LX2- 200c and LX2-nccells were detected by western blot.The proliferation and migration of LX2-200c and LX2-nccellswere measured through CCK-8 assay and wound healing assay respectively. [ Results] The level of miR-200c was low in the wild HSC .The expression of ot-SMA and Vimentin in HSC were enhanced after miR-200c up-regulation. The growth and the migration of HSC was increased by miR-200c up-regulation too. Finally,the level ofcollagen type I (amain component of ECM) expression in LX2-200c cells was significantly higher than that in LX2-nc cells (P = 0.0031 ). [Conclusion] miR-200c involves in liver fibrosis progression through inducing the activation, the growth, the migration and ECM secretion of HSC.
出处
《中山大学学报(医学科学版)》
CAS
CSCD
北大核心
2016年第4期497-501,共5页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金(81201918)
广东省科技计划项目(2012B031800099)