Wang Y, Liang X, Wu S, Murrell G A, Doe W F
Division of Molecular Medicine, John Curtin School of Medical Research, The Australian National University, Canberra, Australia.
Int J Cancer. 2001 Apr 15;92(2):257-62. doi: 10.1002/1097-0215(200102)9999:9999<::aid-ijc1178>3.0.co;2-6.
The role of urokinase-type plasminogen activator receptor (uPAR) in human colon cancer metastasis has not been tested using an antisense approach. In our study, the HCT116 cells, with high metastatic potential were transfected with expression vectors containing a 3' or 5' uPAR cDNA fragment in an antisense (AS) orientation. Transfection of 4 clones was confirmed by DNA hybridization analysis. Receptor-bound endogenous uPA activities of the clones were reduced to 16-68% of controls. The extracellular matrix degradation by the 4 clones was decreased to 33-76%. Two of the clones, 3'-AS7 and 5'-AS, were evaluated in an in vivo assay system of experimental metastasis using athymic mice. Pulmonary metastases were found in 63-78% mice injected with the parent HCT116 or control cells. In mice injected intravenously with the antisense transfected clones, 3'-AS7 and 5'-AS, however, pulmonary metastases were found in only 19% and 9% respectively (p < 0.05). These results provide direct evidence that both 3' and 5'-AS uPAR can inhibit colon cancer invasion and metastasis and may offer the prospect of defining specific targets for gene therapy.
尿激酶型纤溶酶原激活物受体（uPAR）在人类结肠癌转移中的作用尚未通过反义方法进行检测。在我们的研究中，将具有高转移潜能的HCT116细胞用含有反义（AS）方向的3'或5'uPAR cDNA片段的表达载体进行转染。通过DNA杂交分析确认了4个克隆的转染。这些克隆的受体结合内源性uPA活性降低至对照的16 - 68%。4个克隆对细胞外基质的降解降低至33 - 76%。使用无胸腺小鼠在实验性转移的体内测定系统中评估了其中2个克隆，即3'-AS7和5'-AS。在注射亲本HCT116或对照细胞的小鼠中，63 - 78%出现了肺转移。然而，在静脉注射反义转染克隆3'-AS7和5'-AS的小鼠中，肺转移分别仅在19%和9%的小鼠中出现（p < 0.05）。这些结果提供了直接证据，表明3'和5'-AS uPAR均可抑制结肠癌的侵袭和转移，并可能为确定基因治疗的特定靶点提供前景。
