Sequence-specific DNA-binding proteins that bind to the long terminal repeat (LTR) of Moloney leukemia virus in undifferentiated and differentiated mouse embryonal carcinoma (EC) cells were identified by gel retardation assay. The proteins that bind to the CCAAT box were present in both undifferentiated and differentiated EC cells. The amounts and the number of species of the proteins that bind to the enhancer and the GC-rich region were far lower in undifferentiated EC cells than in the differentiated counterparts. These proteins were supposed to be transcriptional activators. Proteins that bind upstream of the enhancer, namely, the -352 to -346 region and the -407 to -404 region, were identified. These proteins were designated the embryonic LTR-binding protein (ELP) and the LTR-binding protein, respectively. The ELP was present only in undifferentiated EC cell lines. The LTR-binding protein was detected in all cell lines tested. The mechanism of suppression of the LTR was investigated by the chloramphenicol acetyltransferase assay. The enhancer and the GC-rich region of the LTR functioned poorly in undifferentiated cells. When eight copies of ELP-binding sequences were inserted upstream of the enhancer region, expression of the chloramphenicol acetyltransferase gene was reduced about threefold in ECA2 cells. From these data, we concluded that two mechanisms, the shortage of activator proteins and the presence of a negative regulatory protein (ELP), are involved in the suppression of the LTR in undifferentiated EC cells.

E‐Cadherin is a member of the cadherin family, which plays a key role in intercellular adhesion in various tumors as well as in normal tissues. Here, we examined the expression of this adhesion molecule in a murine ovarian tumor line OV2944, whose sublines show different degrees of spontaneous metastasis from subcutaneous sites; sublines LM‐1 and LM‐3 exhibit a low metastatic activity but a variant subline HM‐1 has a high metastatic activity. When the expression of E‐cadherin in these cells was examined by immunoblot analysis, the highly metastatic HM‐1 cells was found to express an extremely small amount of this molecule, as compared with a high level of E‐cadherin expression in the weakly metastatic LM‐1 and LM‐3 cells. Northern blot analysis showed that the amount of tanscripts from the E‐cadherin gene is proportional to the amount of proteins detected in these cells. Immunofluorescence staining revealed that cells of the highly metastatic line were heterogeneous, that is, their cultures contained both E‐cadherin‐positive and negative cells. In contrast, cells of the weakly metastatic lines homogeneously expressed E‐cadherin. When the highly metastatic line was subcloned, all the subclones consisted of E‐cadherin‐positive and negative cells. These results suggest that the expression of E‐cadherin gene is not stably controlled in the highly metastatic line.

Nonadherent tissue culture cell lines were established from normal bone marrow of a variety of mouse strains. The lines possessed morphological and histochemical markers of the basophil/mast cell and contained committed stem cells for metachromatic cells. Their derivation from normal marrow and their lack of tumorigenicity despite long-term culture makes these cell lines potentially important for studies of the mechanisms of allergic reactions and inflammation as well as the differentiation pathways involving this subset of hematopoietic cells.

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Natural vitamin E and synthetic vitamin E (dl/‐α‐tocopheryl acetate) were tested for their tnmorigenicity in rodents. Transplantable tumors, at the site of injection, were induced by repeated injections of these compounds in two strains of mice, NFS/N and C57BL/6N × C3H/He Fl, and in a strain of rats, Fischer 344. Natural vitamin E was tumorigenic in both strains of female mice only when injected with soya oil. In contrast, dl‐α‐tocopheryl acetate alone was capable of inducing tumors in Fischer 344 rats. Only one out of 5 male NFS/N mice given dl‐α‐tocopheryl acetate developed a tumor. Therefore, Fischer 344 rats were more susceptible to tumor formation by dl‐α‐tocopheryl acetate than NFS/N mice. dl‐α‐Tocopheryl acetate with soya oil or with palm oil also resulted in the formation of transplantable tumors in NFS/N mice and Fischer 344 rats. There was no difference in the tumor incidence between mice treated with dl‐α‐a‐tocopheryl acetate alone and dl‐α‐tocopheryl acetate plus soya oil or palm oil. However, in rats, the incidence was lower for a group treated with dl‐α‐tocopheryl acetate plus palm oil than for those with dl‐αa‐tocopheryl acetate alone and with dl‐α‐tocopheryl acetate plus soya oil.