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目的观察泮托拉唑对肝癌细胞HepG2迁移和侵袭的影响并探讨其作用机制。方法取对数生长期的HepG2细胞,进行细胞迁移实验和细胞侵袭实验。细胞迁移实验分为正常对照组和泮托拉唑不同浓度组(20、40、80、120、160μmol·L~(-1)),用细胞小室(Transwell)观察泮托拉唑对细胞迁移的影响。细胞侵袭实验分为正常对照组、佛波酯(TPA)组及泮托拉唑(20、40、80、120、160μmol·L~(-1))+TPA(200 nmol·L~(-1))组,用Transwell小室上铺设基质胶观察泮托拉唑对细胞侵袭的影响。采用实时逆转录聚合酶链反应(real time RT-PCR)方法检测细胞中基质金属蛋白酶9(MMP9)mRNA的变化。结果迁移实验表明,与正常对照组比较,泮托拉唑(40、80、120、160μmol·L~(-1))组穿过膜的细胞数明显减少(P<0.01)。侵袭实验表明,TPA组穿过膜的细胞数明显高于正常对照组(P<0.01),泮托拉唑(40、80、120、160 mol·L~(-1))+TPA组穿过膜的细胞数均少于TPA组(P<0.05或P<0.01)。与正常对照组比较,80μmol·L~(-1)泮托拉唑组MMP9 mRNA表达水平下降(P<0.05)。结论泮托拉唑能抑制肝癌细胞HepG2迁移和侵袭,其机制可能与下调MMP9 mRNA的表达有关。
Objective To observe the effect of pantoprazole on the migration and invasion of hepatocellular carcinoma HepG2 cells and to explore its mechanism. Methods HepG2 cells in logarithmic growth phase were used for cell migration assay and cell invasion assay. Cell migration experiments were divided into normal control group and different concentrations of pantoprazole group (20, 40, 80, 120, 160μmol·L -1), with the cell compartment (Transwell) observed pantoprazole cell migration influences. Cell invasion assay was divided into normal control group, phorbol ester (TPA) group and pantoprazole (20,40,80,120,160μmol·L -1) + TPA (200 nmol·L -1 )) Group, using Matrigel on Transwell chamber to observe the effect of pantoprazole on cell invasion. The changes of MMP9 mRNA were detected by real time reverse transcription-polymerase chain reaction (RT-PCR). Results Migration assay showed that the cell number of pantoprazole (40, 80, 120 and 160 μmol·L -1) decreased significantly (P <0.01) compared with the normal control group. The invasion assay showed that the number of cells passing through the membrane in the TPA group was significantly higher than that in the normal control group (P <0.01), and that of the pantoprazole group (40,80,120,160 mol·L -1) + TPA group Membrane cells were less than TPA group (P <0.05 or P <0.01). Compared with the normal control group, the expression of MMP9 mRNA decreased in 80μmol·L -1 pantoprazole group (P <0.05). Conclusion Pantoprazole can inhibit the migration and invasion of HepG2 cells, which may be related to the down-regulation of MMP9 mRNA expression.