Department of General Surgery, College of Clinical Medical Sciences, China Medical University, Shenyang, Liaoning 110004, China.
Chin Med J (Engl). 2012 Jun;125(11):2032-40.
Pancreatic cancer is a devastating disease with the worst mortality rate. Therefore, a rational strategy for future drug development is critical. Genistein is a small, biologically active flavonoid that is found in high amounts in soy. This important compound supports a wide variety of biological activities, but is best known for its ability to inhibit cancer progression.
Transwell chamber assay was performed to determine the effect of genistein on the invasion of the human pancreatic cancer cell line Panc-1 induced by transforming growth factor-β1 (TGF-b1) in the different condition (5 ng/ml 24 hours and 10 ng/ml 48 hours); Reverse transcription-polymerase chain reaction (RT-PCR) was used to estimate the mRNA levels of urinary plasminogen activator (uPA), matrix metallopeptidase 2/9 (MMP-2/9), Smad4, E-Cadherin and Vimentin; Western blotting was used to detect the protein levels of uPA, E-Cadherin, ERK1/2, P38 and P-P38, and the activity of MMP-2/9 protein were detected by gelatin zymography assay method. Cells structure was observed and analyzed by microscopy.
Genistein can inhibit effectively TGF-b1-induced invasion and metastasis in Panc-1 by Transwell assay, which is through regulating the mRNA and protein expression of uPA and MMP2, but not MMP9 by RT-PCR/Western blotting, and is positively correlated with the concentration of genistein. At the same time, genistein also could improve the progress of epithelial-mesenchymal transition (EMT) via morphology observation using light microscopy/transmission electron microscopy (TEM), which is mediated by the down-regulation of E-cadherin and the up-regulation of vimentin.
TGF-b1 mediates EMT process via numerous intracellular signal transduction pathways. The potential molecular mechanisms are all or partly through Smad4-dependent and -independent pathways (p38 MAPK) to regulate the antitumor effect of genistein.
胰腺癌是一种死亡率极高的毁灭性疾病。因此，制定合理的未来药物开发策略至关重要。染料木黄酮是一种小分子、生物活性黄酮类化合物，在大豆中含量很高。这种重要的化合物支持广泛的生物活性，但最著名的是其抑制癌症进展的能力。
采用 Transwell 室试验，在不同条件下（5ng/ml24 小时和 10ng/ml48 小时）观察染料木黄酮对转化生长因子-β1（TGF-β1）诱导的人胰腺癌细胞系 Panc-1 侵袭的影响；采用逆转录-聚合酶链反应（RT-PCR）估计尿纤溶酶原激活物（uPA）、基质金属蛋白酶 2/9（MMP-2/9）、Smad4、E-钙粘蛋白和波形蛋白的 mRNA 水平；采用 Western blot 检测 uPA、E-钙粘蛋白、ERK1/2、P38 和 P-P38 的蛋白水平，采用明胶酶谱法检测 MMP-2/9 蛋白的活性。通过显微镜观察和分析细胞结构。
染料木黄酮通过 Transwell 试验有效抑制 TGF-β1 诱导的 Panc-1 侵袭和转移，这是通过调节 uPA 和 MMP2 的 mRNA 和蛋白表达实现的，但不是通过 RT-PCR/Western blot 检测到的 MMP9，且与染料木黄酮的浓度呈正相关。同时，染料木黄酮还可以通过光镜/透射电镜（TEM）观察细胞形态，改善上皮-间质转化（EMT）的进程，这是通过下调 E-钙粘蛋白和上调波形蛋白介导的。
TGF-β1 通过多种细胞内信号转导途径介导 EMT 过程。潜在的分子机制均部分或全部通过 Smad4 依赖性和非依赖性途径（p38MAPK）来调节染料木黄酮的抗肿瘤作用。
