伏立诺他抑制胰腺癌PANC-1细胞增殖及其血管生成拟态的相关研究

Study of the inhibition effect of SAHA on cell proliferation and vasculogenic mimicry in pancreatic cancer PANC-1 cells

目的研究伏立诺他(SAHA)对人胰腺癌PANC-1细胞体外增殖、迁移及其细胞凋亡的影响,观察SAHA对胰腺癌PANC-1细胞主导的血管生成拟态的作用以及对PANC-1细胞中基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)表达水平的影响。方法倒置显微镜下观察不同浓度的SAHA对PANC-1细胞增殖的作用。MTT比色法检测SAHA对PANC-1细胞的毒性作用。行细胞划痕试验观察SAHA对PANC-1细胞迁移的作用。流式细胞仪分析SAHA对胰腺癌PANC-1细胞凋亡的影响。利用细胞三维培养技术观察经SAHA诱导24 h后的胰腺癌PANC-1细胞体外类血管结构形成情况。Western blotting技术检测SAHA处理后的胰腺癌PANC-1细胞中MMP-2的表达水平。结果 SAHA能显著抑制人胰腺癌PANC-1细胞的生长,且呈明显的剂量依赖性关系。与对照组相比,SAHA组均能抑制PANC-1细胞迁移(均P<0.01)。SAHA可诱导人胰腺癌PANC-1细胞发生细胞凋亡。体外三维培养显示,SAHA组较对照组均能抑制PANC-1细胞主导的血管生成拟态(均P<0.05)。随着SAHA浓度的递增,PANC-1细胞中MMP-2的表达水平呈现下降趋势(P<0.05)。结论组蛋白去乙酰化酶抑制剂SAHA能有效抑制人胰腺癌PANC-1细胞体外增殖、迁移并诱导其发生细胞凋亡,SAHA下调MMP-2的表达可能是SAHA抗胰腺癌血管生成拟态的机制之一。

Objective To investigate the effects of suberoylanilide hydroxamic acid(SAHA), an inhibitor of histone deacetylase, on the cell proliferation, cell migration and apoptosis of pancreatic cancer PANC-1 cells. To observe the changes of vasculogenic mimicry dominated by pancreatic cancer PANC-1 cells and the expression level of matrix metalloproteinase-2(MMP-2) in PANC-1 cells which treated with different concentrations of SAHA. Methods Inverted microscope was used to observe the effect of a series of concentrations of SAHA in cell proliferation of pancreatic cancer PANC-1 cells. MTT colormetric assay was performed to measure the inhibitory effect of different concentration of SAHA treatment for 24 h on human pancreatic cancer PANC-1 cells. Cell scratch test and was used to measure the effect of SAHA on the migration abilities of PANC-1 cells, and then, we use flow cytometry to analyze the effect of SAHA on cell apoptosis. PANC-1 cells were treated with the different concentration of SAHA for 24 h, vasculogenic-like networks were formed in three-dimensional culture in vitro, then we observe the changes in the structures of vasculogenic-like network. Western blotting was used to study the expression level of MMP-2 in pancreatic cancer PANC-1 cells with the different concentration of SAHA treatment. Results SAHA could significantly inhibit the proliferation of pancreatic cancer PANC-1 cells and showed the dose-dependent relationships. Cell scratch test showed that SAHA could effectively inhibit the migration of PANC-1 cells compared to the control group(P<0.01). Flow cytometry showed that SAHA could obviously induce the apoptosis of pancreatic cancer PANC-1 cells. Three-dimensional culture in vitro showed that SAHA could significantly inhibit vasculogenic mimicry dominated by pancreatic cancer PANC-1 cells(P<0.05). Western blotting showed that with the increase concentration of SAHA, the expression level of MMP-2 in PANC-1 cells of pancreatic cancer decreased(P<0.05). Conclusion SAHA can effectively inhibit the proliferation and migration of PANC-1 cells leads to cell apoptosis. Meanwhile, the down-regulation of MMP-2’ s expression caused by SAHA may be one of the mechanisms of it’ s anti-tumor mimicry in pancreatic cancer.