Morgan John C, Majors John E, Galileo Deni S
Department of Neurology, Medical College of Georgia, Augusta, GA 30912, USA.
Clin Exp Metastasis. 2005;22(2):167-75. doi: 10.1007/s10585-005-6930-4.
The cellular proto-oncogene c-src is thought to be involved in formation, progression, and metastasis of a variety of tumor cell types, although its exact role during tumor cell genesis is not well defined. v-src, the viral oncogene counterpart of c-src, causes metastatic sarcomas, hemorrhagic disease, and hemangiosarcomas in chicken embryos and, thus, can be used as a constitutively activated form of src for experimentally-induced tumorigenesis. Here, we used retroviral vectors to express wild-type v-src or SH2 or SH3 domain-deleted forms (DeltaSH2 or DeltaSH3) to determine if different pathogenic effects resulted. Vectors were injected into early chick embryo midbrain ventricles and embryos were sacrificed at various ages up to embryonic day (E) 18. Retroviral expression of all forms of v-src resulted in transformation of pial connective tissue cells into large, rounded abnormal-appearing cells. Surprisingly, all forms of v-src were lethal. The v-src retrovirus was lethal and killed most embryos by E15 with the development of hemangiosarcomas over the injection site between E10-E12. The DeltaSH3 retrovirus was the most deadly, killing most embryos by E12, however, it never resulted in hemangiosarcoma formation. The DeltaSH2 retrovirus injected embryos survived longer than v-src or DeltaSH3 embryos, and some of these embryos also developed large hemangiosarcomas over the injection site between E13 and E18. These results demonstrate that the src SH2 domain is required to be fully lethal, whereas the presence of the SH3 domain attenuated lethality. Furthermore, the formation of hemangiosarcomas absolutely required the presence of the src SH3 domain and to some extent required the SH2 domain. This implicates distinct and opposite roles for SH2 and SH3 domains of src and their cellular binding partners in tumorigenesis and hemorrhagic disease.
细胞原癌基因c-src被认为与多种肿瘤细胞类型的形成、进展和转移有关,尽管其在肿瘤细胞发生过程中的具体作用尚不清楚。v-src是c-src的病毒癌基因对应物,可在鸡胚中引发转移性肉瘤、出血性疾病和血管肉瘤,因此可作为src的组成型激活形式用于实验诱导的肿瘤发生。在此,我们使用逆转录病毒载体表达野生型v-src或SH2或SH3结构域缺失形式(DeltaSH2或DeltaSH3),以确定是否会产生不同的致病效应。将载体注射到早期鸡胚中脑脑室,并在胚胎发育至第18天(E18)的不同年龄段处死胚胎。所有形式的v-src的逆转录病毒表达均导致软脑膜结缔组织细胞转化为大的、圆形的异常细胞。令人惊讶的是,所有形式的v-src都是致命的。v-src逆转录病毒是致命的,在E15时杀死了大多数胚胎,在E10 - E12期间注射部位出现血管肉瘤。DeltaSH3逆转录病毒最为致命,在E12时杀死了大多数胚胎,然而,它从未导致血管肉瘤形成。注射DeltaSH2逆转录病毒的胚胎比v-src或DeltaSH3胚胎存活时间更长,其中一些胚胎在E13和E18之间注射部位也出现了大的血管肉瘤。这些结果表明,src的SH2结构域是完全致死所必需的,而SH3结构域的存在减弱了致死性。此外,血管肉瘤的形成绝对需要src的SH3结构域的存在,并且在一定程度上需要SH2结构域。这暗示了src的SH2和SH3结构域及其细胞结合伴侣在肿瘤发生和出血性疾病中具有截然不同且相反的作用。