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S Estrade

Publications and source records attributed to S Estrade.

32 records · Page 2Linked to original sources

Simian virus 40-Chinese hamster kidney cell interaction. IV. Enhanced virus replication in infected cells upon treatment with mitomycin C.

Chinese hamster kidney cells are semi-permissive to simian virus 40 (SV40). Exposure to mitomycin C (MC) of Chinese hamster kidney cells infected with SV40 DNA enhanced the yield of infectious virus 10- to 100-fold. This stimulation occurred whether the treatment was performed before or after infection. A simultaneous increase in the number of V antigen-synthesizing cells and virus-producing cells, as well as the virus burst size, was observed upon MC pretreatment, whereas the proportion of T antigen-synthesizing cells remained unchanged. MC pretreatment clearly stimulated virus DNA replication in SV40 virus-infected cells. Cells treated with MC exhibited an unbalanced growth pattern, with continuing protein synthesis in the absence of cell division and a markedly reduced ability to replicate the cellular DNA. These results suggest that MC enhances the permissiveness of Chinese hamster kidney cells by inducing the synthesis of a specific cellular factor(s) required for SV40 reproduction in Chinese hamster kidney cells.

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Simian virus 40-chinese hamster kidney cell interaction. II. The semipermissivity of the cell system.

Throughout in vitro passages, Chinese hamster kidney (CHK) cells progressively lost susceptibility to SV 40 virus infection while remaining continuously susceptible to viral DNA infection. Upon infection with SV 40 virus or viral DNA, the CHK cell line supported viral DNA and virus replication at a low level. SV 40 transformed CHK cell lines spontaneously produced small amounts of viral DNA and virions. The percentage of virus-producing cells was low. Various clones derived from each of these lines behaved as the parental cell population, leading to the conclusion that each CHK cell, whether transformed or not with SV 40, is potentially permissive for this virus.

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Simian virus 40-chinese hamster kidney cell interaction. I. Relationship of chromosome changes to transformation.

After approximately 20 in vitro passages, Chinese hamster kidney (CHK) cell cultures transformed upon exposure to different strains of SV 40 can show a diploid modal chromosome number of 22 with chromosome counts exclusively or essentially in the diploid range (20-25). In primary culture and at the 5th-7th subculture, tumors produced in nude mice by such cells can display a diploid modal value of 22 chromosomes. Numerical chromosome variations and karyotype abnormalities observed in CHK cells transformed upon infection with SV 40, are apparently indistinguishable from those that can be observed in uninfected CHK cells undergoing "spontaneous"-transformation. These results indicate that polyploidization is not a necessary step either in the process of transformation or in that of tumorigenic conversion with SV 40.

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Activation of the viral genome in Simian Virus 40-transformed nonpermissive cells by permissive cell extracts.

Simian virus 40-transformed nonpermissive cells in which neither infectious viral DNA nor virions had previously been detected reproducibly yield low levels of infectious simian virus-40 DNA after treatment with extracts of normal permissive cells. Virions were obtained in one out of 37 experiments. Activation of the viral genome is detected only with normal permissive cell extracts and only in simian virus 40-transformed cell lines that are rescuable by heterokaryon formation or by treatment with chemicals. The activating factor(s) is insensitive to deoxyribonuclease and ribonuclease, but is sensitive to heat at 56 degrees and to proteolytic enzymes.

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Use of monkey-mouse hybrid cells for the study of the cellular regualtion of interferon production and action.

Four clones of a somatic monkey-mouse hybrid cell line were studied. One of these clones produced both mouse and monkey interferon, while the three others produced only mouse interferon. All four were, however, sensitive to both mouse and monkey (or human) interferon. The karyotype analysis of these four hybrid clones and the parental cell lines enabled us to locate the possible genetic site governing the synthesis of monkey interferon on a small subtelocentric monkey chromosome. The genetic site responsible for the synthesis of the antiviral protein is located on a different (monkey or mouse) chromosome.

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Cellular genes possibly involved in the transformation process of the human melanoma cell line XP44 RO (Mel).

P44 Ro (Mel) is a human malignant melanoma cell line derived from a testicular metastasis in a DNA repair deficient, xeroderma pigmentosum patient. This line harbors a N-ras gene mutated in codon 61. To investigate other cellular genes possibly contributing to the expression of its transformed phenotype, four XP44 revertant cell lines were isolated by different selection procedures and the association of the level of expression of various oncogenes (including N-ras) and tumor suppressor genes with the selection for the revertant phenotype was determined. The revertants exhibited a significant but variable degree of phenotypic reversion, according to the selective pressure to which they were submitted, and a phenotypic stability dependent on their constant maintenance in selective medium. Back-revertant lines were isolated by culturing revertant lines in control medium for several weeks. The comparison between parental, revertant and back-revertant cells has revealed that, beyond the mutation in codon 61 of N-ras, two groups of genes appear to be also implicated in the transformation process of XP44 RO (Mel) cells: one group, comprising pim A, trk, Rb and p53, whose expression is independent of the cell selection conditions; the other group, comprising Ha-ras, N-ras, neu 1, fos and met H, whose expression is more or less dependent upon such conditions. The myc gene is apparently not involved in this phenomenon. These results, besides strengthening the concept that carcinogenesis is a multigenic process, suggest that diverse mechanisms can lead to the transformed phenotype, but that these mechanisms might have some pathway(s) in common.

3T3 Cells↗

Polyamine content and oncogene expression in hepatoma cells in culture during methionine deprivation and refeeding.

From an hepatocarcinoma cell line (LFCL.2A), unable to grow in a culture medium in which methionine was replaced by L-homocysteine, we had previously isolated revertant clones presenting a low growth rate, a loss of tumorigenicity and an inhibition of transcription of three oncogenes: c-Ki-ras, c-Ha-ras and c-myc. Here we showed that long-term deprivation of methionine led to a depletion of spermine, while putrescine and spermidine contents remained unchanged. When the revertant cells were shifted in a medium containing methionine, the oncogene transcription (except the p53 gene) started very rapidly in parallel with an increase in the putrescine content. By contrast, spermidine and spermine contents decreased during the first hours but were not significantly different from control values after numerous subcultures in methionine-containing medium.

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