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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 HMGB1 gene on invasion and migration of human hepatocellular carcinoma cell line HepG2 and its related mechanisms. Methods siRNA targeting HMGB1 was synthesized by RNAi technique and transfected into HepG2 cells. Transmembrane migration assay and planar scratch healing assay were used to detect the invasion and migration of cells before and after transfection. The expressions of MMP-2, MMP-9 and MMP-9 were detected by RT-PCR and Western blot respectively. Intercellular adhesion molecule 1 (ICAM-1) and tissue inhibitor of metalloproteinase 2 (TIMP-2) mRNA and protein expression. Results HepG2 cells were transfected with 40 nmol / L HMGB1-siRNA for 24 h, and both the planar and transmembrane migration activities were significantly inhibited compared with the control group. The expressions of MMP-2, MMP-9, ICAM -1 mRNA expression was significantly down-regulated, while TIMP-2 mRNA expression was significantly up-regulated (P <0.05). Conclusion HMGB1 can promote the invasion and migration of hepatocellular carcinoma cells, which is related to the regulation of the expression of MMP-2, MMP-9, ICAM-1 and TIMP-2 by HMGB-1.