Cavanaugh P G, Nicolson G L
Department of Tumor Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Cancer Res. 1989 Jul 15;49(14):3928-33.
The ability of malignant cells to respond to growth factor(s) present in or secreted by a distant target organ may be important in tumor metastasis. We used metastatic cell lines and clones of the rat 13762NF mammary adenocarcinoma that show reproducible spontaneous metastatic behavior from the mammary fat pad to regional lymph nodes and lung sites. Whereas poorly lung metastatic MTPa and MTC cells did not grow in response to lung-conditioned medium, highly lung-metastatic MTLn3 cells responded and grew rapidly in lung-conditioned medium. The major growth-promoting factor for MTLn3 cells from porcine and rat lung-conditioned media was purified by using hydroxylapatite affinity and anion exchange chromatography, chromatofocusing, size exclusion chromatography, and preparative native gel electrophoresis. The activity in each of the purification fractions was measured by determining their ability to increase the number of MTLn3 cells in serum-deprived culture. The major component that differentially stimulated the growth of highly metastatic MTLn3 cells was a glycoprotein of Mr approximately 66,000. Under reducing conditions, its apparent Mr was approximately 72,000. This lung-derived mitogen was stable at pH 4.0-9.0, possessed a pI of 6.9-7.0, and preferentially promoted the growth of lung-metastasizing tumor lines over their poorly lung-metastasizing counterparts in rat 13762NF mammary adenocarcinoma and murine B16 melanoma tumor systems. The activity of porcine lung-derived growth factor was not affected by pretreatment with antisera to porcine insulin, human granulocyte-macrophage colony-stimulating factor, human platelet-derived growth factor, or murine epidermal growth factor. It was inactivated by reduction with dithiothreitol or exposure to high temperature (95 degrees C). The results suggest that specific organ-derived growth factors are important in metastatic colonization and organ growth of particular malignant cells.
恶性细胞对远处靶器官中存在或分泌的生长因子作出反应的能力,可能在肿瘤转移中起重要作用。我们使用了大鼠13762NF乳腺腺癌的转移细胞系和克隆,这些细胞系和克隆表现出从乳腺脂肪垫到区域淋巴结和肺部的可重复的自发转移行为。肺转移能力差的MTPa和MTC细胞对肺条件培养基无生长反应,而肺转移能力强的MTLn3细胞对肺条件培养基有反应并快速生长。通过使用羟基磷灰石亲和层析、阴离子交换层析、色谱聚焦、尺寸排阻层析和制备性天然凝胶电泳,从猪和大鼠肺条件培养基中纯化出了促进MTLn3细胞生长的主要生长因子。通过测定各纯化组分增加血清饥饿培养中MTLn3细胞数量的能力,来检测其活性。差异刺激高转移性MTLn3细胞生长的主要成分是一种分子量约为66,000的糖蛋白。在还原条件下,其表观分子量约为72,000。这种源自肺的有丝分裂原在pH 4.0 - 9.0时稳定,pI为6.9 - 7.0,在大鼠13762NF乳腺腺癌和小鼠B16黑色素瘤肿瘤系统中,优先促进肺转移肿瘤系的生长,而不是肺转移能力差的对应肿瘤系。猪肺源性生长因子的活性不受抗猪胰岛素、人粒细胞 - 巨噬细胞集落刺激因子、人血小板源性生长因子或鼠表皮生长因子抗血清预处理的影响。用二硫苏糖醇还原或高温(95℃)处理可使其失活。结果表明,特定器官源性生长因子在特定恶性细胞的转移定植和器官生长中起重要作用。