摘要
目的探讨高迁移率族蛋白B1(HMGB1)基因对人肝癌细胞株HepG2侵袭迁移的影响及相关机制。方法应用RNA干扰(RNAi)技术合成针对HMGB1的siRNA并转染HepG2。跨膜迁移实验、平面划痕愈合实验检测转染前后细胞侵袭迁移能力的变化;逆转录PCR(RT-PCR)及Western blot法分别测定基质金属蛋白酶2(MMP-2)、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mRNA和蛋白表达。结果用40 nmol/L的HMGB1-siRNA转染HepG2细胞24 h,与对照组相比,细胞的平面迁移活性及跨膜迁移活性均受到明显抑制;转染后MMP-2、MMP-9、ICAM-1的mRNA表达明显下调,而TIMP-2的mRNA表达明显上调(P<0.05)。结论 HMGB1可促进肝癌细胞的侵袭迁移,该作用与HMGB-1调控MMP-2、MMP-9、ICAM-1和TIMP-2的表达有关。
Objective To investigate the effect of high mobility group-box protein 1 （HMGBI）-siRNA on invasion and migration of human hepatoma cell line HepG2, and further to explore its mechanism. Methods HMGB]-siRNA was synthesized with RNA interference and transferred into HepG2 cells to down-regulate the expression of HMGBI. The invasion and migration activities were assayed by TranswellTM assay and monolayer wounding healing assay. The levels of matrix metallo- proteinase 2 （MMP-2）, MMP-9, intercellular adhesion molecule 1 （ICAM-I） 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 HMGBI-siRNA for 24 h. Results The migration and invasion abilities of HepG2 cells were inhibited by 40 nmol/L HMGBt-siRNA markedly, Compared with the control group, MMP-2, MMP-9, ICAM-1 were down-regulated, TIMP-2 was up-regulated significantly by HMGBI-siRNA （ P 〈 0.05）. Conclusion HMGBI-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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2013年第11期1159-1162,共4页
Chinese Journal of Cellular and Molecular Immunology
