水飞蓟宾通过Smad信号通路抑制TGF-β诱导的结直肠癌HT-29细胞中的MMP-2和MMP-9

Z. Zare, Tina Nayerpour dizaj, Armaghan Lohrasbi, Zakieh Sadat Sheikhalishahi, Amirhooman Asadi, Mana Zakeri, F. Hosseinabadi, Omid Abazari, M. Abbasi, Parisa Khanicheragh
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引用次数: 12

摘要

背景:肿瘤细胞转移是导致结直肠癌(CRC)患者死亡的主要原因。转化生长因子-β (TGF-β)诱导的基质金属蛋白酶(MMPs)在转移过程中至关重要。水飞蓟宾是从水飞蓟中提取的一种天然化合物,对癌细胞具有抗肿瘤活性。在本研究中,我们评估水飞蓟宾对TGF-β诱导的人HT-29 CRC细胞株MMP-2和MMP-9的影响及其潜在机制。方法:在HT-29细胞系上进行体外实验。HT-29细胞系在RPMI1640中培养,在不含和存在不同浓度水飞蓟宾(10、25、50和100 μM)的情况下暴露于TGFβ (5 ng/ml)中。MTT法测定水飞蓟宾对HT-29细胞活力的影响。实时聚合酶链反应(real-time PCR)检测MMP-2和MMP-9 mRNA的相对表达量。Western blotting检测MMP-2、MMP- 9蛋白表达及Smad2磷酸化水平。结果:水飞蓟宾对HT-29细胞株24小时细胞活力有一定的抑制作用,且呈剂量依赖性。与对照组相比,TGF-β增加了MMP-2、MMP-9和磷酸化Smad2的mRNA和蛋白表达。水飞蓟宾药理抑制显著阻断TGF-β诱导的MMP-2和MMP-9 mRNA和蛋白表达及Smad2磷酸化。结论:水飞蓟宾降低HT-29癌细胞的细胞活力呈剂量依赖性。水飞蓟宾还能抑制TGF-β刺激的HT-29细胞中MMP-2和MMP-9的表达,可能是通过Smad2信号通路介导的。
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Silibinin Inhibits TGF-β-induced MMP-2 and MMP-9 Through Smad Signaling Pathway in Colorectal Cancer HT-29 Cells
Background: Metastasis of cancer cells is the primary responsible for death in patients with colorectal cancer (CRC). Transforming growth factor-β (TGF-β)-induced matrix metalloproteinases (MMPs) are essential for the metastasis process. Silibinin is a natural compound extracted from the Silybum marianum that exhibits anti-neoplastic activity in cancer cell lines. In this study, we evaluated the effects of silibinin on MMP-2 and MMP-9 induced by TGF-β in human HT-29 CRC cell line and the potential mechanism underlying the effects. Methods: The present in vitro study was done on the HT-29 cell line. The HT-29 cell line was cultured in RPMI1640 and exposed to TGFβ (5 ng/ml) in the absence and presence of different concentrations of silibinin (10, 25, 50, and 100 μM). The effect of silibinin on HT-29 cell viability was measured with the MTT assay. A real-time polymerase chain reaction (Real-Time PCR) determined the relative mRNA expression of MMP-2 and MMP-9. Western blotting was employed to examine MMP-2 and MMP 9 protein expression and Smad2 phosphorylation. Results: Silibinin inhibits cell viability of HT-29 cell line at 24 hours in a dose-dependent manner. TGF-β increased the mRNA and protein expression of MMP-2, MMP-9, and phosphorylated Smad2 compared to controls. Pharmacological inhibition with silibinin markedly blocked TGF-β–induced MMP-2 and MMP-9 mRNA and protein expression and Smad2 phosphorylation. Conclusion: Silibinin decreased the cell viability of HT-29 cancer cells in a dose-dependent manner. Silibinin also inhibited TGF-β-stimulated MMP-2 and MMP-9 expression in HT-29 cells, possibly mediated with the Smad2 signaling pathway.
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