Increased cell-substrate adhesion accompanies conditional reversion to the normal phenotype in ras-oncogene-transformed NIH-3T3 cells.
Increased cell-substrate adhesion accompanies conditional reversion to the normal phenotype in ras-oncogene-transformed NIH-3T3 cells.
复制标题
在 ras 癌基因转化的 NIH-3T3 细胞中，细胞与基质粘附力的增加伴随着向正常表型的有条件恢复。
DOI：
10.1006/excr.1994.1280
复制
发表时间：
1994
影响因子：
3.7
通讯作者：
Varani,J
中科院分区：
文献类型：
--
作者：
Shumaker,DK;Sklar,MD;Prochownik,EV;Varani,J
我们最近报道(1991,Mol。在ras转化的NIH-3T3细胞中，通过myc反义序列消耗c-myc蛋白可以逆转这些细胞的一些恶性特征。这些包括转化形态，在软琼脂中生长，以及在胸腺小鼠中形成肿瘤的能力。在本研究中，我们检测了相同的细胞的体外粘附行为。与ras转化的NIH-3T3细胞相比，通过反义转染耗尽c-myc蛋白的细胞对纤维连接蛋白包被的培养皿的附着没有变化，但在附着后对胰蛋白酶/ edta介导的基质释放的抵抗力大大增加。在相关研究中，检测了细胞的纤维连接蛋白生物合成和细胞表面纤维连接蛋白。与ras转化的NIH-3T3相比，c-myc反义转染细胞中纤维连接蛋白的生物合成没有总体变化。然而，免疫荧光染色显示与反义表达c-myc的转染物相关的表面纤维连接蛋白数量增加。综上所述，这些数据表明，在ras转化的NIH-3T3细胞中，通过选择性地消耗c-myc蛋白而有条件地重新获得非恶性表型与细胞-底物粘附的增加有关。这反过来又与表面纤维连接蛋白的增加有关。
We recently reported (1991,Mol. Cell Biol. 11, 3699-3710) that depletion of c-myc protein by myc antisense sequences in ras-transformed NIH-3T3 cells reverses several of the malignant characteristics of these cells. These include transformed morphology, growth in soft agar, and ability to form tumors in athymic mice. In the present study we examined the same cells forin vitroadhesive behavior. Cells depleted of c-myc protein by antisense transfection showed no change in attachment to fibronectin-coated dishes as compared to ras-transformed NIH-3T3 cells but had greatly increased resistance to trypsin/EDTA-mediated release from the substratum after attachment. In concomitant studies, the cells were examined for fibronectin biosynthesis and cell surface fibronectin. There was no overall change in fibronectin biosynthesis in the c-myc antisense transfected cells as compared to the ras-transformed NIH-3T3. However, immunofluorescence staining revealed increased amount of surface fibronectin associated with the antisense c-myc-expressing transfectants. Taken together, these data indicate that the conditional reacquisition of the nonmalignant phenotype in ras-transformed NIH-3T3 cells by selected depletion of c-myc protein is associated with an increase in cell-substrate adhesion. This, in turn, is associated with an increase in surface fibronectin.