Spontaneous metastasis of rat liver epithelial cells transformed with v-raf and v-raf/v-myc: association with different phenotypic properties.

Invasion & metastasis Pub Date : 1997-01-01
H C Bisgaard, A R Mackay, D E Gomez, P T Ton, S S Thorgeirsson, U P Thorgeirsson
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Abstract

Cloned v-raf, v-raf/v-myc, and spontaneously transformed rat liver epithelial (RLE) cell lines were examined for meastatic capability in nude mice, using the LacZ gene as a marker for quantitation of micrometastases. Six cloned lines (R3611-T lines) derived from nude mouse xenografts of the v-raf transformed R3611-3 cells displayed variable metastatic capabilities. Three of six subcutaneously inoculated R3611-TlacZ lines produced spontaneous lung metastasis in nude mice. One of the lines, R3611-T2lacZ was highly efficient at metastatic conversion and produced more lung colonies than a faster growing v-raf/v-myc-transformed RJ2-14lacZ line. The spontaneously transformed RLElacZ line (C4T) was nonmetastatic, although it produced larger subcutaneous tumors than the metastatic R3611-T2lacZ line. Metastatic conversion correlated with upregulation of urokinase-type plasminogen activator receptor RNA expression and downregulation of plasminogen activator inhibitor-1, collagen alpha1 (I), and cytokeratin 14 (K14) RNA expression. These findings indicate that proteolytic activities associated with plasminogen activation play a role in the metastatic development in this model. Decreased production of extracellular proteins and cytoskeletal changes associated with lack of K14 expression are also likely to have contributed to the metastatic conversion of the RLE transformants.

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v-raf和v-raf/v-myc转化大鼠肝上皮细胞的自发转移:与不同表型特性的关联
克隆的v-raf、v-raf/v-myc和自发转化的大鼠肝上皮(RLE)细胞系在裸鼠身上的转移能力被检测,使用LacZ基因作为微转移的定量标记。从v-raf转化的R3611-3细胞的裸鼠异种移植中获得的6个克隆系(R3611-T系)显示出不同的转移能力。6个皮下接种的R3611-TlacZ细胞系中有3个在裸鼠中产生自发性肺转移。其中一株R3611-T2lacZ在转移转化方面效率很高,比生长更快的v-raf/v-myc转化的RJ2-14lacZ株产生更多的肺菌落。自发转化的RLElacZ系(C4T)虽然比转移的R3611-T2lacZ系产生更大的皮下肿瘤,但没有转移性。转移转化与尿激酶型纤溶酶原激活物受体RNA表达上调和纤溶酶原激活物抑制剂-1、胶原α 1 (I)和细胞角蛋白14 (K14) RNA表达下调相关。这些发现表明,与纤溶酶原激活相关的蛋白水解活性在该模型的转移发展中起作用。细胞外蛋白的减少和与K14表达缺乏相关的细胞骨架变化也可能导致RLE转化子的转移性转化。
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