Dose-effects of 8-methoxypsoralen and long wave UV-light in 3T3 cells: evaluation of a phototoxic index

Summary 3T3 cells were cultured until confluency and treated with various doses of 8-methoxypsoralen followed by long wave UV light irradiation. The inhibition of³H-thymidine incorporation was dose-dependent for both, psoralen and light. A phototoxic index (PTI) was established demonstrating that a constant correlation between psoralen and UVA light exists for the photoinactivation in living cells.Supported by Deutsche Forschungsgemeinschaft.We thank Miss Anne Schröder for her skillful assistance.     

Conversion of psoralen DNA monoadducts in E. coli to interstrand DNA cross links by near UV light (320-360 nm): inability of angelicin to form cross links, in vivo.

Both angelicin and psoralen monoadducts formed in vivo in E. coli by near UV light produce lethal and mutagenic effects. However psoralen monoadducts are converted to cross links by higher doses of UV; angelicin monoadducts are not. The relevance of these results to psoralen photosensitization is discussed.     

Conversion of psoralen DNA monoadducts in E. coli to interstrand DNA cross links by near UV light (320–360 nm): Inability of angelicin to form cross links,in vivo

Summary Both angelicin and psoralen monoadducts formed in vivo in E. coli by near UV light produce lethal and mutagenic effects. However psoralen monoadducts are converted to cross links by higher doses of UV; angelicin monoadducts are not. The relevance of these results to psoralen photosensitization is discussed.Supported by the National Research Council of Canada.     

Prolongation of the survival of skin grafts in mice by PUVA treatment

The combined application of psoralen and UVA radiation to skin grafts induced a prolongation of the survival time of the grafts in mice. This was observed using the H-Y barrier, an allogeneic barrier without H-2 disparities, and a strong H-2 incompatible barrier. The effect is probably due to a reduction of antigen-presenting cells, or to other, unknown mechanisms.     

Prolongation of the survival of skin grafts in mice by PUVA treatment

Summary The combined application of psoralen and UVA radiation to skin grafts induced a prolongation of the survival time of the grafts in mice. This was observed using the H-Y barrier, an allogeneic barrier without H-2 disparities, and a strong H-2 incompatible barrier. The effect is probably due to a reduction of antigen-presenting cells, or to other, unknown mechanisms.     

In situ demonstration of the activity of 3 beta-hydroxysteroid dehydrogenase on steroid hormone secreting cells within the paraaortic lymph nodes of golden hamster

The in situ activity of 3 beta-hydroxysteroid dehydrogenase was detected in clustered cells which showed steroid-producing morphology, within the capsule of the paraaortic lymph node. In light and electron microscopic studies, the positive reaction products were detected on intracapsular cell clusters. This result indicates that these unique cells may have a steroid secreting function within the lymph nodes.     

Presence of specific binding sites for [3H]sarcophytol-A in cultured human skin fibroblasts

The presence of specific binding sites for [3H]sarcophytol-A in human skin fibroblasts was examined using biochemical and morphological methods. The displacement studies clearly revealed that high (KD = 31.0 nM) and low (KD = 6.05 microM) affinity sites were present in the intact cells. Moreover, autoradiographic studies using light microscopy revealed that the specific binding sites may exist in both the cytoplasm and the nuclei.     

Effet de l'ATP sur l'incorporation d'un DNA-H3 exogène dans les cellules HeLa en culture

Summary Cultures of HeLa cells treated with ATP show an increased uptake of DNA-H³ for a short period of treatment. In the light ofGropp's findings on pinocytosis activation, it is tempting to suppose that the stimulation of DNA-uptake by ATP is mediated through an activity of the latter on pinocytosis.     

Presence of specific binding sites for [3H]sarcophytol-A in cultured human skin fibroblasts

Summary The presence of specific binding sites for [³H]sarcophytol-A in human skin fibroblasts was examined using biochemical and morphological methods. The displacement studies clearly revealed that high (K_D=31.0 nM) and low (K_D=6.05 M) affinity sites were present in the intact cells. Moreover, autoradiographic studies using light microscopy revealed that the specific binding sites may exist in boththe cytoplasm and the nuclei.     

Activation of Janus kinase/signal transducer and activator of transcription 3 pathway by growth hormone-releasing hormone

Growth hormone-releasing hormone (GHRH) can act as a potent growth factor in various cancers. The mitogenic activity of this neuropeptide is exerted through binding to the pituitary type receptors (GHRH-R) or their splice variants (SV). In the present work, we studied whether this hormone can activate the JAK2/STAT3 pathway which plays a crucial role in cancer cell proliferation and is also linked to carcinogenesis. We transfected HeLa human cervical cancer cells, which are not sensitive to GHRH analogs with the pGHRH-R. Transfected cells responded to the GHRH or its antagonist with an increase or a decrease in proliferation, respectively. These results were confirmed by the expression of proliferating cell nuclear antigen. We then showed that these effects are linked to the activation of the JAK2/STAT3 pathway. Our work demonstrates the activation of JAK/STAT3 pathway by GHRH and sheds further light to the mechanisms of the antitumorogenic action of GHRH antagonists.     

<Emphasis Type="Italic">XPF</Emphasis> knockout via CRISPR/Cas9 reveals that ERCC1 is retained in the cytoplasm without its heterodimer partner XPF

The XPF/ERCC1 heterodimeric complex is essentially involved in nucleotide excision repair (NER), interstrand crosslink (ICL), and double-strand break repair. Defects in XPF lead to severe diseases like xeroderma pigmentosum (XP). Up until now, XP-F patient cells have been utilized for functional analyses. Due to the multiple roles of the XPF/ERCC1 complex, these patient cells retain at least one full-length allele and residual repair capabilities. Despite the essential function of the XPF/ERCC1 complex for the human organism, we successfully generated a viable immortalised human XPF knockout cell line with complete loss of XPF using the CRISPR/Cas9 technique in fetal lung fibroblasts (MRC5Vi cells). These cells showed a markedly increased sensitivity to UVC, cisplatin, and psoralen activated by UVA as well as reduced repair capabilities for NER and ICL repair as assessed by reporter gene assays. Using the newly generated knockout cells, we could show that human XPF is markedly involved in homologous recombination repair (HRR) but dispensable for non-homologous end-joining (NHEJ). Notably, ERCC1 was not detectable in the nucleus of the XPF knockout cells indicating the necessity of a functional XPF/ERCC1 heterodimer to allow ERCC1 to enter the nucleus. Overexpression of wild-type XPF could reverse this effect as well as the repair deficiencies.     

肝癌细胞系Hep3B中CD133+细胞亚群的分离及其特性的研究

目的研究CD133在人肝癌细胞系Hep3B中的表达以及CD133+细胞的体外增殖、自我更新及体内成瘤能力,初步探讨肝癌中CD133+细胞亚群的干细胞特性。方法流式细胞仪检测未分选的Hep3B细胞中CD133+细胞表达情况;免疫磁珠分选技术纯化CD133+肿瘤细胞;MTT法检测CD133+细胞体外增殖能力;无血清培养纯化...     

Serum-induced stimulation of snRNA synthesis in mouse 3T3 fibroblasts

Small nuclear RNAs (snRNAs) from quiescent and serum-stimulated 3T3 cultures, labeled with [3H]uridine [( 3H]U), were electrophoresed in polyacrylamide-urea slab gels and revealed by staining with ethidium bromide and by fluorography. Judged by labeling with [3H]U, synthesis of 7S and U1-U6 RNAs was very low or absent in quiescent cultures. The serum-induced transition of 3T3 cells from a resting to a growing state was accompanied by an early, apparently sequential stimulation of snRNA synthesis; stimulated synthesis of 7S, U1, U2, U3, U4 and U6 RNAs coincided in time with serum-induced stimulation of 45S pre-ribosomal RNA (pre-rRNA) and heterogeneous nuclear RNA (hnRNA) synthesis.     

Insulin-like effect of dichloroacetic acid on hexose transport in Swiss 3T3 cells

Summary Hexose transport in Swiss 3T3 cells was increased by treatment with dichloroacetic acid as well as by treatment with insulin. Neither extra-nor intracellular Ca²⁺ was found to be involved in their stimulatory action. On the other hand, the removal of intracellular Mg²⁺ resulted in a loss of the stimulation. These results suggest that dichloroacetic acid stimulates the hexose transport in Mg²⁺-dependent manner, similar to that of insulin.This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture, and the Ministry of Health and Welfare of Japan.     

Serum-induced stimulation of snRNA synthesis in mouse 3T3 fibroblasts

Summary Small nuclear RNAs (snRNAs) from quiescent and serum-stimulated 3T3 cultures, labeled with [³H]uridine ([³H]U), were electrophoresed in polyacrylamide-urea slab gels and revealed by staining with ethidium bromide and by fluorography, Judged by labeling with [³H]U, synthesis of 7S and U1-U6 RNAs was very low or absent in quiescent cultures. The serum-induced transition of 3T3 cells from a resting to a growing state was accompanied by an early, apparently sequential stimulation of snRNA synthesis; stimulated synthesis of 7S, U1, U2, U3, U4 and U6 RNAs coincided in time with serum-induced stimulation of 45S pre-ribosomal RNA (pre-rRNA) and heterogeneous nuclear RNA (hnRNA) synthesis.     

Light evoked release of radioactivity from rabbit retinas preloaded with (3H)-GABA

Summary Light flashes evoke an increased release of radioactivity in vitro from rabbit retinas preloaded with (³H)-GABA in vivo. Constant light does not affect the release. No light evoked release can be demonstrated from the glia. Pentobarbitone and AOAA depress the evoked release. The results are consistent with GABA being a retinal neurotransmitter, most likely in a class of amacrines.This work was supported by grants from the Swedish Medical Research Council (project 04X2321), The Faculty of Medicine, University of Lund, Royal Physiographic Society.