Nie Jinlan, Huang Quanfang, Tan Shimei, Lin Xing
Department of Pharmacy, Guangxi Medical University, Nanning 530021, China.
Department of Pharmacy, First Affiliated Hospital, Guangxi University of Chinese Medicine, Nanning 530023, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Nov;32(11):1491-1494.
Objective To investigate the effect of Raf kinase inhibitor protein (RKIP) over-expression on the proliferation of LX-2 human hepatic stellate cells. Methods Recombinant plasmid pcDNA3.1-RKIP was transfected into LX-2 cells. G418 was used to screen and culture stably infected cells. MTT assay and colony formation assay were used to examine the effect of RKIP over-expression on cell proliferation and colony formation, respectively. Western blotting was performed to assess the expressions of RKIP, α-smooth muscle actin (α-SMA), type 1 collagen (Col1) and matrix metalloproteinase 1(MMP-1) and MMP-2 as well as extracellular signal-regulated kinases/mitogen-activated protein kinase (ERK/MAPK) signaling pathway-related proteins. Results Compared with the control cells, RKIP over-expression significantly inhibited LX-2 cell proliferation and colony formation, and reduced the protein expressions of Col1, α-SMA, MMP-1 and MMP-2. Moreover, RKIP over-expression remarkably inhibited the phosphorylation of ERK/MAPK. Conclusion Over-expressed RKIP inhibits LX-2 cell proliferation and the mechanism is related to the inhibition of ERK/MAPK signaling pathway.
目的 探讨Raf激酶抑制蛋白（RKIP）过表达对LX-2人肝星状细胞增殖的影响。方法 将重组质粒pcDNA3.1-RKIP转染至LX-2细胞。用G418筛选并培养稳定感染的细胞。分别采用MTT法和集落形成试验检测RKIP过表达对细胞增殖和集落形成的影响。进行蛋白质免疫印迹法检测RKIP、α-平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原（Col1）、基质金属蛋白酶1（MMP-1）和MMP-2以及细胞外信号调节激酶/丝裂原活化蛋白激酶（ERK/MAPK）信号通路相关蛋白的表达。结果 与对照细胞相比，RKIP过表达显著抑制LX-2细胞增殖和集落形成，并降低Col1、α-SMA、MMP-1和MMP-2的蛋白表达。此外，RKIP过表达显著抑制ERK/MAPK的磷酸化。结论 RKIP过表达抑制LX-2细胞增殖，其机制与抑制ERK/MAPK信号通路有关。
