Department of Pharmacy, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, Guangxi, China.
Life Sciences Institute and Pharmaceutical College, Guangxi Medical University, Nanning, Guangxi, China.
J Cell Biochem. 2019 Apr;120(4):6168-6177. doi: 10.1002/jcb.27904. Epub 2018 Oct 10.
The purpose of this study was to investigate the effect of Raf kinase inhibitor protein (RKIP) on the growth, apoptosis, invasion, and metastasis of human hepatic stellate cell line (LX-2). A recombinant plasmid (pcDNA3.1-RKIP) or RKIP-targeting small interfering RNA (siRNA) vector (siRNA-RKIP) was transfected into LX-2 cells to interfere with the RKIP expression. The results demonstrated that increased RKIP expression significantly reduced cell viability, clonogenic growth, and invasion. Further, it promoted cell apoptosis and induced cell cycle arrest in the G1 phase. Overexpression of RKIP led to inactivation of LX-2 cells, as evidenced by the decrease in the expression levels of collagen I and α-smooth muscle actin (α-SMA). In addition, increased RKIP expression significantly reduced the phosphorylation of Raf/extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK), the transcriptional activity of nuclear factor-κB (NF-κB), and the levels of matrix metalloproteinases-1 and -2. In conclusion, these findings clearly demonstrate that RKIP inhibits LX-2 cell growth, metastasis, and activation, primarily by downregulating the ERK/MAPK and NF-κB signaling pathways.
本研究旨在探讨 Raf 激酶抑制剂蛋白（RKIP）对人肝星状细胞系（LX-2）生长、凋亡、侵袭和转移的影响。将重组质粒（pcDNA3.1-RKIP）或 RKIP 靶向小干扰 RNA（siRNA）载体（siRNA-RKIP）转染入 LX-2 细胞，以干扰 RKIP 的表达。结果表明，RKIP 表达增加显著降低细胞活力、集落形成能力和侵袭能力。此外，它促进细胞凋亡并诱导细胞周期停滞在 G1 期。RKIP 的过表达导致 LX-2 细胞失活，这表现在胶原 I 和α-平滑肌肌动蛋白（α-SMA）的表达水平降低。此外，RKIP 表达增加显著降低 Raf/细胞外信号调节激酶（ERK）/丝裂原活化蛋白激酶（MAPK）、核因子-κB（NF-κB）转录活性以及基质金属蛋白酶-1 和 -2 的水平。总之，这些发现清楚地表明，RKIP 通过下调 ERK/MAPK 和 NF-κB 信号通路抑制 LX-2 细胞的生长、转移和激活。
