Kurer M A
Department of Surgery, York District Hospital, Wigginton Road, York, YO31 8HE, UK.
Mol Biol Rep. 2007 Dec;34(4):221-4. doi: 10.1007/s11033-006-9036-4. Epub 2006 Dec 19.
Patients with solo tumour malignancy are at higher risk of developing venous thromboembolism. When prophylactic anticoagulation (and in particular heparin) is used during cancer therapy however, patients appear to have a prolonged survival. Tumours express large quantities of procoagulant molecules, which predispose patients to these conditions. Tissue Factor (TF) is an important example, which may have a role in the biology of malignant disease. Intra-tumour vessel coagulation however is not a common phenomenon. Our hypothesis is that cancer cells produce anticoagulant molecules, which may prevent intra-tumour vessel auto-coagulation. Our results show that one such factor--Tissue Factor Pathway Inhibitor (TFPI-1) is expressed by a number of different cancer cells.
Seven human cancer cell lines were studied: three breast, two colorectal and two pancreatic. Cells were maintained in cell culture, and at 90% confluence protein and RNA were extracted. RNA integrity was confirmed using an RNA integrity gel and RNA purity determined by spectrophotometry. Reverse transcription polymerase chain reaction (RT-PCR) was used for TFPI-1 mRNA detection and immunoblotting used for TFPI-1 protein detection.
Six cell lines (two breast, two colorectal, and two pancreatic) expressed the TFPI-1 gene. Gene function was confirmed by detection of TFPI-1 protein expression in these cell lines.
TFPI-1 is expressed by breast cancer and other cancer cell lines maintained in cell culture. This has not been previously reported. Functional expression of TFPI-1 by cancer cells suggests that it has an important role in cancer biology. Further experiments are required to establish its function.
孤立肿瘤恶性肿瘤患者发生静脉血栓栓塞的风险较高。然而,在癌症治疗期间使用预防性抗凝治疗(尤其是肝素)时,患者的生存期似乎会延长。肿瘤表达大量促凝分子,这使患者易患这些疾病。组织因子(TF)就是一个重要例子,它可能在恶性疾病生物学中发挥作用。然而,肿瘤内血管凝血并非常见现象。我们的假设是癌细胞产生抗凝分子,这可能会阻止肿瘤内血管的自动凝血。我们的结果表明,一种这样的因子——组织因子途径抑制剂(TFPI - 1)由多种不同癌细胞表达。
研究了七种人类癌细胞系:三种乳腺癌细胞系、两种结肠癌细胞系和两种胰腺癌细胞系。细胞在细胞培养中维持生长,当达到90%汇合度时提取蛋白质和RNA。使用RNA完整性凝胶确认RNA完整性,并通过分光光度法测定RNA纯度。逆转录聚合酶链反应(RT - PCR)用于检测TFPI - 1 mRNA,免疫印迹用于检测TFPI - 1蛋白。
六个细胞系(两个乳腺癌细胞系、两个结肠癌细胞系和两个胰腺癌细胞系)表达TFPI - 1基因。通过检测这些细胞系中的TFPI - 1蛋白表达证实了基因功能。
TFPI - 1在细胞培养中维持的乳腺癌和其他癌细胞系中表达。此前尚未有此报道。癌细胞对TFPI - 1的功能性表达表明它在癌症生物学中具有重要作用。需要进一步实验来确定其功能。