Dabbous M K, El-Torky M, Haney L, Sobhy N, Brinkley S B
Exp Mol Pathol. 1983 Feb;38(1):1-21. doi: 10.1016/0014-4800(83)90094-1.
Primary and secondary cultures of VX-2 carcinoma produced high levels of collagenase activity in both active and latent forms in serum-free media. These cultures appeared morphologically heterogeneous in phase-contrast microscopy and revealed the presence of mainly three distinct forms: epithelial-like cells (E cells), fibroblast-like cells (F cells), and large rounded-flat cells which may represent a subclass of the F cells. Cell separation techniques such as brief dispase treatment, Percoll gradient centrifugation, thimerosal treatment, and rabbit serum were used to obtain predominantly one form or the other. The E cells never formed a monolayer but rather grew as limited size clusters of intimately associated cells with large nuclei and often appeared multinucleated. These cells were difficult to maintain in culture or serially passed more than a few times. The F cells, rare in early cultures but having the highest growth potential, appeared in various morphological forms ranging from spindle- to stellate-shaped cells. The cells in their third passage were capable of producing palpable tumors, similar in light and electron microscopic studies to the original tumor from which they were derived, when injected intramuscularly into recipient rabbits and produced specific collagenase activity in active and latent forms in serum-free media. Ultrastructural studies suggested that the E cells were of epithelial origin whereas the F cells were similar to stromal fibroblasts. Cytogenetic studies demonstrated that almost all of the E cells showed both numerical and structural chromosomal changes in a modal number of 54 chromosomes. On the other hand, the major cell population of the F cells resembled normal rabbit fibroblasts; both contained a normal diploid (2n = 44). However, few cells (4-6%) in the F-cell population were hyperdiploid with a modal chromosome number of 54. These cells may represent inadvertent contaminating E cells and account for the apparent limited turmorigenicity observed in early F-cell cultures. The data suggested that the E cells were of tumor origin whereas the majority of the F-cell population appeared to be of host origin. Furthermore, it is suggested that the E cells stimulate tumor-associated stromal cells to produce elevated levels of collagenolytic activity and contribute to collagen degradation during tumor invasion.
VX-2 癌的原代和传代培养物在无血清培养基中产生了高水平的活性和潜伏形式的胶原酶活性。这些培养物在相差显微镜下形态上呈现异质性，主要显示出三种不同的形式：上皮样细胞（E 细胞）、成纤维细胞样细胞（F 细胞）以及大的圆形扁平细胞，后者可能代表 F 细胞的一个亚类。采用诸如短暂的 dispase 处理、Percoll 梯度离心、硫柳汞处理和兔血清等细胞分离技术来主要获得一种或另一种细胞形式。E 细胞从未形成单层，而是以紧密相连的细胞组成的有限大小的簇生长，细胞核大，且常出现多核现象。这些细胞在培养中难以维持，或传代次数超过几次就很困难。F 细胞在早期培养物中很少见，但具有最高的生长潜力，呈现出从纺锤形到星状等各种形态。第三代的这些细胞在无血清培养基中能产生活性和潜伏形式的特异性胶原酶活性，当肌肉注射到受体兔体内时，在光镜和电镜研究中产生的可触及肿瘤与它们所源自的原始肿瘤相似。超微结构研究表明，E 细胞起源于上皮，而 F 细胞类似于基质成纤维细胞。细胞遗传学研究表明，几乎所有的 E 细胞在 54 条染色体的众数上都显示出数量和结构上的染色体变化。另一方面，F 细胞的主要细胞群体类似于正常兔成纤维细胞；两者都含有正常的二倍体（2n = 44）。然而，F 细胞群体中少数细胞（4 - 6%）是超二倍体，众数染色体数为 54。这些细胞可能代表无意中污染的 E 细胞，并解释了在早期 F 细胞培养物中观察到的明显有限的致瘤性。数据表明，E 细胞起源于肿瘤，而大多数 F 细胞群体似乎起源于宿主。此外，有人认为 E 细胞刺激肿瘤相关的基质细胞产生升高水平的胶原olytic 活性，并在肿瘤侵袭过程中促进胶原蛋白降解。
