Synthesis and evaluation of biological activities of vibsanin A analogs

Vibsanin A is an 11-membered vibsane diterpenoid and is reported to induce myeloid cell differentiation via activation of protein kinase C (PKC) without tumor-promoting activity. Therefore, vibsanin A is thought to be an attractive compound for acute myeloid leukemia (AML) therapy. In this study, we synthesized vibsanin A analogs and compared the activity of these compounds for PKC activation and myeloid cell differentiation. We found that the hydroxymethyl group in vibsanin A is an important substituent to induce differentiation of AML cells. Collectively, our results showed the biochemical features of vibsanin A and provided new insights into the development of new antileukemic drugs.     

Regulation of N-glycosylation and secretion of Isthmin-1 by its C-mannosylation

BackgroundC-mannosylation is a type of protein glycosylation. Human Isthmin-1 (ISM1) is a 52-kDa secreted protein with a thrombospondin type 1 repeat (TSR) domain, containing two consensus C-mannosylation sequences at Trp²²³ and Trp²²⁶. In this study, we sought to examine the role of C-mannosylation in the secretion of ISM1.MethodsWe established and cultured an ISM1-overexpressing HT1080 cell line and purified recombinant ISM1 for analysis from the conditioned medium by LC-MS/MS. Subcellular localization of ISM1 was observed by confocal fluorescence microscopy.ResultsWe found that ISM1 is C-mannosylated at Trp²²³ and Trp²²⁶ in the TSR domain. To determine the functions of the C-mannosylation of ISM1, we established a C-mannosylation-defective mutant ISM1-overexpressing HT1080 cell line and measured its secretion of ISM1. The secretion of ISM1 decreased significantly in this mutant ISM1-overexpressing line compared with wild-type cells. Furthermore, ISM1 was N-glycosylated only in these C-mannosylation-defective cells.ConclusionsISM1 is C-mannosylated in its TSR domain, and the status of the C-mannosylation of ISM1 affects its N-glycosylation.General significanceThe C-mannosylation of ISM1 regulates its N-glycosylation status.     

Cell-dependent regulation of vasculogenic mimicry by carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1)

Vasculogenic mimicry (VM) promotes tumor migration, metastasis, and invasion in various types of cancer, but the relationship between VM and these phenotypes remains undefined. In this study, we examined carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1) as a novel target of VM. We found that ectopic expression of CEACAM1 in HT1080 human fibrosarcoma cells suppressed the formation of a VM-like network. Further, cell migration and proliferation were abated by the introduction of CEACAM1 into HT1080 cells. Conversely, knockout (KO) of the CEACAM1 gene in SK-MEL-28 melanoma cells, which normally express high levels of CEACAM1, inhibited formation of a VM-like network, which was covered on reintroduction of CEACAM1. These results suggest that CEACAM1 differentially regulates formation of the VM-like network between cancer cell types and implicate CEACAM1 as a novel therapeutic target in malignant cancer.     

Mapping the dominant wound healing and soft tissue regeneration QTL in MRL × CAST

We have used a mouse ear punch model and the QTL (quantitative trait loci) mapping technique to identify genes that are responsible for soft tissue regeneration. In the early studies, we have identified several QTL and have shown that the inheritance of ear healing was additive in one cross (MRL × SJL), and recessive in another cross (DBA × 129). Because CAST mice are genetically distinct and have a different genetic background, CAST would facilitate the identification of common and novel QTL when crossed with common inbred lines. We made a cross between super healer MRL and poor healer CAST and collected ear punch phenotype and marker genotype data from F₂. Ear punch healing exhibited a dominant mode of inheritance in this cross. There were three main QTL on Chromosomes 4, 9, and 17, and two suggestive QTL on Chromosomes 1 (new) and 7. Taken together, these QTL accounted for about 29% of total F₂ variance of MRL × CAST. Compared with another study using the same cross, we found a totally different set of QTL. Two QTL interactions were identified by a full QTL model: Chromosomes 4 × 17 and 9 × 17; the latter reached to a statistical level at p < 0.05. These interactions explained about 4% of the F₂ phenotypic variance. We conclude that soft tissue regeneration is controlled by multiple genes and locus vs. locus interactions. This work was supported by Assistance Award No. DAMD17-99-1-9571. The U.S. Army Medical Research Acquisition Activity, Fort Detrick, MD, is the awarding and administering acquisition office. The information contained in this publication does not necessarily reflect the position or policy of the U.S. Government and no official endorsement should be inferred.     

Intraspecific polymorphism of pollen sculpture inParis tetraphylla A. Gray (Liliaceae)

Paris tetraphylla has a greater diversity of pollen exine sculpture than was previously thought. On the basis of 357 plants from 52 localities of Japan, we distinguished seven sculpture types that are tentatively referred to here as G₁, G₂, G₃, R₁, R₂, R₃ and S. G₁, G₂ and G₃ have gemmate sculpture in which the gemmae are characteristically large, medium and small in size, respectively; likewise, R₁, R₂ and R₃, have coarsely to finely ruglate sculpture; and S, scrobiculate sculpture. In general, any one of the seven types always or predominantly occurs in each plant, thus allowing us to make comparisons among plants and further among local populations. We thus found that G₂ and G₃, respectively, are the most and the second most widespread and occur in plants of 41 and 22 respectively, of the localities examined, although their frequencies differ with locality. Generally R₃ occurs in plants distributed at higher latitudes, while R₂ is rather widespread and common to plants growing on high mountains. G₁ and R₁ are restricted to plants occurring south of Ibaraki Pref. and north of Gunma Pref., respectively. Such localized distributional patterns of some types may reflect the reproductive nature and the history of geographical isolation inParis tetraphylla over the Japanese Islands.