Wang Chao, Tang Chaofeng, Chang Xiaowei, Li Zhaoyu
School of Graduates, General Hospital, Ningxia Medical University, Yinchuan 750004, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Nov;29(11):1159-62.
To investigate the effect of high mobility group-box protein 1 (HMGB1)-siRNA on invasion and migration of human hepatoma cell line HepG2, and further to explore its mechanism.
HMGB1-siRNA was synthesized with RNA interference and transferred into HepG2 cells to down-regulate the expression of HMGB1. The invasion and migration activities were assayed by Transwell™ assay and monolayer wounding healing assay. The levels of matrix metalloproteinase 2 (MMP-2), MMP-9, intercellular adhesion molecule 1 (ICAM-1) and tissue inhibitor of MMP-2 (TIMP-2) were detected by RT-PCR and Western blotting in HepG2 cells after treatment with 40 nmol/L HMGB1-siRNA for 24 h.
The migration and invasion abilities of HepG2 cells were inhibited by 40 nmol/L HMGB1-siRNA markedly. Compared with the control group, MMP-2, MMP-9, ICAM-1 were down-regulated, TIMP-2 was up-regulated significantly by HMGB1-siRNA (P<0.05).
HMGB1-siRNA can inhibit the invasion and migration abilities of human hepatoma cells by down-regulating the expressions of MMP-2, MMP-9, ICAM-1 and up-regulating the expression of TIMP-2.
探讨高迁移率族蛋白B1（HMGB1）-小干扰RNA（siRNA）对人肝癌细胞系HepG2侵袭和迁移能力的影响，并进一步探究其作用机制。
采用RNA干扰技术合成HMGB1-siRNA，并将其转染至HepG2细胞中以下调HMGB1的表达。通过Transwell™实验和单层划痕愈合实验检测细胞的侵袭和迁移活性。用40 nmol/L HMGB1-siRNA处理HepG2细胞24小时后，采用逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹法检测基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）、细胞间黏附分子1（ICAM-1）和基质金属蛋白酶组织抑制因子2（TIMP-2）的水平。
40 nmol/L HMGB1-siRNA显著抑制了HepG2细胞的迁移和侵袭能力。与对照组相比，HMGB1-siRNA使MMP-2、MMP-9、ICAM-1表达下调，TIMP-2表达显著上调（P<0.05）。
HMGB1-siRNA可通过下调MMP-2、MMP-9、ICAM-1的表达及上调TIMP-2的表达来抑制人肝癌细胞的侵袭和迁移能力。
