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目的观察高转移性肝癌细胞系HCC9204细胞转染抑制性κB基因(IκB-α)后核因子κB(NF-κB)和基质金属蛋白酶-9(MMP-9)表达的改变以及细胞浸润转移能力的改变。方法采用脂质体法将IκB-α基因转染HCC9204细胞,应用Western-blot及RT-PCR法检测MMP-9和NF-κB的表达;电泳迁移率分析(EMSA)测定NF-κB活性;免疫组化法检NF-κBp65、MMP-9的表达;激光扫描共聚焦显微镜(LSCM)观察MMP-9细胞定位以及细胞形态;基底膜侵袭实验检测肿瘤细胞的浸润和转移。结果HCC9204细胞转染PcDNA3-IκB-α后,细胞内NF-κB和MMP-9mRNA表达以及NF-κB活性下降;NF-κB、MMP-9表达具有明显的正相关;MMP-9蛋白表达位置和细胞形态改变;细胞侵袭转移能力明显下降。结论NF-κB表达水平与HCC9204的浸润转移能力有关;NF-κB活性抑制后细胞浸润转移能力下降,MMP-9表达下降可能起主要作用。
Objective To investigate the expression of nuclear factor κB (NF-κB) and matrix metalloproteinase-9 (MMP-9) and the ability of cell invasion and metastasis in transfected HCC9204 cell line with high metastatic hepatocellular carcinoma cell line HCC9204 change. Methods The IκB-α gene was transfected into HCC9204 cells by liposome. The expression of MMP-9 and NF-κB was detected by Western-blot and RT-PCR. The activity of NF-κB was measured by electrophoretic mobility shift assay (EMSA) The expression of NF-κBp65 and MMP-9 was detected by histochemical method. The localization and cell morphology of MMP-9 cells were observed by laser scanning confocal microscopy (LSCM). The invasion and metastasis of tumor cells were detected by invasion assay. Results After pcDNA3-IκB-α was transfected into HCC9204 cells, the expression of NF-κB and MMP-9 mRNA and the activity of NF-κB were decreased. The expressions of NF-κB and MMP-9 were positively correlated. Cell morphology changes; cell invasion and metastasis decreased significantly. Conclusions The expression of NF-κB is related to the invasion and metastasis of HCC9204. The decreased ability of infiltration and metastasis of NF-κB and the decrease of MMP-9 expression may play a major role.