Cytochalasin D promotes pulmonary metastasis of B16 melanoma through expression of tissue factor
Cytochalasin D promotes pulmonary metastasis of B16 melanoma through expression of tissue factor
复制标题
细胞松弛素D通过组织因子表达促进B16黑色素瘤肺转移
DOI：
10.3892/or.2013.2423
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发表时间：
2013-07-01
期刊：
影响因子：
4.2
通讯作者：
Lin, Ying-Ying
中科院分区：
文献类型：
--
作者：
Huang, Feng-Ying;Mei, Wen-Li;Lin, Ying-Ying
细胞松弛素D（CytD）作用于肌动蛋白，这是一种在真核细胞中普遍存在的蛋白质。以往的研究主要集中在CytD的抗肿瘤作用上。我们之前意外发现CytD可促进B16黑色素瘤细胞的肺转移，因此，在本研究中我们探究了可能的潜在机制。将B16黑色素瘤细胞与CytD及其他试剂共培养，并用于建立肺转移模型。在这个B16黑色素瘤转移模型中，发现经CytD处理的小鼠肺转移和肺重量显著增加，而通过慢病毒表达的组织因子（TF）RNA干扰几乎完全抑制了这种情况。Northern印迹、Western印迹以及实时逆转录聚合酶链反应（RT-PCR）分析结果显示，在经CytD处理的B16细胞中，TF的表达显著上调，但受到TF RNA干扰的显著抑制。此外，在经CytD处理的转移肺组织以及与CytD和因子VIIa（FVIIa）共培养的B16细胞中，还发现丝裂原活化蛋白激酶p38的上调和磷酸化，但单独用CytD、二甲基亚砜或FVIIa培养的细胞中未出现这种情况。这些结果表明，CytD刺激B16黑色素瘤细胞中TF的表达，通过与FVIIa结合激活凝血依赖性和非依赖性途径，最终促进肺转移。TF干扰是预防B16黑色素瘤转移的一种潜在方法。
Cytochalasin D (CytD) targets actin, a ubiquitous protein in eukaryotic cells. Previous studies have focused mainly on the antitumor effects of CytD. We previously found CytD to promote lung metastasis in B16 melanoma cells, which we had not anticipated, and, therefore, in the present study we investigated the possible underlying mechanisms. B16 melanoma cells were co-cultured with CytD and other agents and used to establish a lung metastatic model. In this B16 melanoma metastatic model, significantly increased lung metastasis and lung weight were found in CytD-treated mice, which was almost completely suppressed by tissue factor (TF) RNA interference expressed via lentivirus. The results of northern and western blot, and real-time RT-PCR analysis showed that the expression of TF was significantly upregulated in B16 cells treated with CytD but was significantly inhibited by TF RNA interference. In addition, upregulation and phosphorylation of mitogen-activated protein kinase p38 were also found in the metastatic lung tissues treated with CytD and in the B16 cells co-cultured with CytD and factor Vila (FVIIa), but not in cells cultured with CytD, dimethyl sulfoxide or FVIIa alone. These results indicate that CytD stimulates the expression of TF in B16 melanoma cells, activating both coagulation-dependent and -independent pathways via binding to FVIIa, eventually promoting lung metastasis. TF interference is a potential approach to the prevention of B16 melanoma metastasis.