研究白介素6(IL-6)在过氧化氢诱导的肺泡上皮细胞凋亡中的作用及其机制。方法：以人Ⅱ型肺泡上皮细胞A549细胞株为实验对象，MTT法检测IL-6对A549细胞增殖活性的影响，TUNEL检测法检测IL-6对A549细胞凋亡率的影响，Western blot观察caspase-3和caspase-9蛋白量的变化。结果：IL-6能够增加A549细胞的增殖活性，减少其凋亡。与模型组相比，IL-6干预组的caspase-3和caspase-9的蛋白表达量明显降低,有统计学意义。结论：IL-6能够抑制过氧化氢诱导的肺泡上皮细胞凋亡，并且与caspase-3和caspase-9蛋白的表达量下降有关。提示IL-6对肺泡上皮细胞的保护作用可能是通过抑制细胞凋亡相关蛋白来实现的。
ObejectiveTo establish the lenvatinib-resistant model of hepatocellular carcinoma (HCC) in vitro in order to study the effect of disulfiram combined with copper ions on the proliferation and apoptosis of lenvatinib-resistant Huh7 cells.MethodsThe half-inhibitory concentration (IC50) of Huh7 cells treated with lenvatinib was detected by the CCK8 method, and the resistance of Huh7 cells to lenvatinib was induced by increasing concentration method. The cell treatment of each group: The Huh7 cells cultured with common culture medium and with 10 μmol/L lenvatinb in the control group and lenvatinib sensitive group, respectively; The lenvatinib-resistant Huh7 cells cultured with 10 μmol/L lenvatinb, with 10 μmol/L lenvatinib+1 μmol/L disulfiram+0.5 μmol/L copper ions, and with 10 μmol/L lenvatinib+10 μmol/L Akt inhibitor MK2206 in the lenvatinib-resistant group, combined drugs group, and inhibitor group, respectively. The cell survival and apoptosis rates were detected by the CCK8 and flow cytometry, respectively. The Western blot method was used to detect the expressions of phospho-protein kinase B (p-Akt), cysteine aspartic acid specific protease (caspase-9), and B-cell lymphoma-2 (Bcl-2) proteins. The Transwell assay was used to detect the cell invasiveness.ResultsThe lenvatinib-resistant cell line was successfully established. The concentrations of 10, 20, 40, 60, 80 and 160 μmol/L leventinib were applied to the Huh7 cells for 48 h, and the IC50 was calculated to be 12.35 μmol/L. The cell survival rate of the lenvatinib-resistant group was significantly higher than that of the lenvatinib sensitive group (P<0.01), which in the combined drugs group was significantly lower than that of the lenvatinib-resistant group (P<0.01), which in the inhibitor group was significantly lower than that of the lenvatinib-resistant group (P<0.05). The apoptosis rate of the combined drugs group was significantly higher than that of the lenvatinib-resistant group (P<0.01). Compared with the lenvatinib-resistant group, the expression of p-Akt protein was significantly decreased (P<0.01) and caspase-9 expression was significantly increased (P<0.01) in the combined drugs group, while the difference of Bcl-2 protein between the lenvatinib-resistant group and the combined drugs group was not statistically significant (P>0.05). The invasive cells were more in the combined drugs group as compared with the lenvatinib-resistant group (P<0.01). ConclusionThe combination of disulfiram and copper ion could increase the sensitivity of lenvatinib to lenvatinib-resistant Huh7 cells, the cell inhibition rate is significantly increased, it’s mechanism might be related to the inhibiting p-Akt expression and promoting caspase-9 expression.