GENETIC TAGGING OF TUMOR-CELLS WITH RETROVIRUS VECTORS - CLONAL ANALYSIS OF TUMOR-GROWTH AND METASTASIS INVIVO
GENETIC TAGGING OF TUMOR-CELLS WITH RETROVIRUS VECTORS - CLONAL ANALYSIS OF TUMOR-GROWTH AND METASTASIS INVIVO
复制标题
DOI：
10.1128/mcb.8.8.3143
复制
发表时间：
1988-08-01
影响因子：
5.3
通讯作者：
KERBEL, RS
中科院分区：
文献类型：
--
作者：
KORCZAK, B;ROBSON, IB;KERBEL, RS
使用逆转录病毒载体感染将大量独特的遗传标记引入肿瘤细胞群中，目的是分析转移性肿瘤与非转移性肿瘤在体内渐进生长过程中克隆组成的比较变化。使用的细胞系是 SP1（一种非转移性非整倍体小鼠乳腺癌）和 SP1HU9L（SP1 的转移性变体）。细胞用.DELTA.e.DELTA.pMoTN感染，这是一种复制缺陷型逆转录病毒载体，其具有显性选择性neo基因和残缺的长末端重复序列。以4次的频率获得G418r集落。 10-3。许多克隆的 Southern 印迹分析提供了一个或两个原病毒 DNA 拷贝的随机和可遗传整合的证据。通过皮下注射来自3.6倍的混合混合物的105个细胞来分析原发性肿瘤生长的克隆进化以及自发转移和原发性肿瘤之间的谱系关系的性质。将 102 个 G418r SP1HU9L 或 104 个 G418r SP1 集落注入同系 CBA/J 小鼠中。最引人注目的发现是晚期原发性肿瘤的相对克隆同质性。尽管注射了数百或数千个带有独特标记的克隆，但它们总是由少量（少于10个）不同的克隆组成。就转移性 SP1HU9L 细胞而言，这些“优势”克隆的性质因肿瘤而异。对许多肺转移瘤的分析表明，其中一部分来自显性的原发肿瘤克隆，并由一种（有时是两种）不同的祖细胞组成。在一些动物中，所有肺转移瘤都源自共同的祖细胞克隆，而在其他动物中，每个转移结节都有不同的祖细胞。结果显示如下。 (i)逆转录病毒载体感染可用于将大量独特且稳定的克隆标记引入肿瘤细胞群中。 (ii) 非常有限数量的克隆的后代在晚期原发性肿瘤中占主导地位。 (iii) 乳腺癌转移是单克隆或双克隆来源的。讨论了结果的意义。
Retrovirus vector infection was used to introduce large numbers of unique genetic markers into tumor cell populations for the purpose of analyzing comparative changes in the clonal composition of metastatic versus that of nonmetastatic tumors during their progressive growth in vivo. The cell lines used were SP1, a nonmetastatic, aneuploid mouse mammary adenocarcinoma, and SP1HU9L, a metastatic variant of SP1. Cells were infected with .DELTA.e.DELTA.pMoTN, a replication-defective retrovirus vector which possesses the dominant selectable neo gene and crippled long terminal repeats. G418r colonies were obtained at a frequency of 4 .times. 10-3. Southern blot analysis of a number of clones provided evidence of random and heritable integration of one or two copies of the proviral DNA. Clonal evolution of primary tumor growth and the nature of lineage relationships among spontaneous metastases and primary tumors were analyzed by subcutaneously injecting 105 cells from a pooled mixture of 3.6 .times. 102 G418r SP1HU9L or 104 G418r SP1 colonies into syngeneic CBA/J mice. The most striking finding was the relative clonal homogeneity of advanced primary tumors; they invariably consisted of a small number (less than 10) of distinct clones depsite the fact that hundreds or thousands of uniquely marked clones had been injected. In the case of the metastatic SP1HU9L cells, the nature of these "dominant" clones varied from one tumor to another. Analysis of a number of lung metastases revealed that a proportion of them were derived from dominant primary tumor clones and were composed of one, and sometimes two, distinct progenitors. In some animals, all the lung metastases were derived from a common progenitor clone, whereas in others, each metastatic nodule had a different progenitor. The results show the following. (i) Retrovirus vector infection can be used to introduce large numbers of unique and stable clonal markers into tumor cell populations. (ii) The progeny of a very limited number of clones dominate in advanced primary tumors. (iii) Mammary carcinoma metastases are of mono- or biclonal origin. The significance of the results is discussed.