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1.
Immortalization of human aortic smooth muscle cells with origin-minus simian virus 40 DNA. 总被引:2,自引:0,他引:2
N Murahashi Y Sasaguri M Ohuchida N Kakita M Morimatsu 《Biotechnology and applied biochemistry》1992,16(2):152-160
We established two cell lines of human smooth muscle cells (SMC) by transfection of cells from the aortic intima and aortic media with origin-minus simian virus 40 (ori-minus SV40) DNA. Ori-minus SV40 DNA very efficiently immortalized human smooth muscle cells in culture. Proteins that these cell lines produced included type I, III, IV, and V collagens, fibronectin, and human matrix metalloproteinases (MMP)-1 (tissue collagenase), -2 ("type IV collagenase"), and -3 (stromelysin). The protein production in these cell lines generally mimicked that of normal SMC, but the immortalization stimulated the cell line of medial SMC to produce excessive MMP-2 and to secrete MMP-9 (92-kDa gelatinase). However, since these cell lines did not show a fully malignant phenotype, we concluded that, in addition to the degradation of extracellular matrix macromolecules, including basement membrane components by MMP-2, -3, and/or -9, some additional factors must be involved for the malignancy of fully transformed cells and that these immortalized human aortic SMC, which share many characteristics with normal SMC, will prove useful to study the role(s) of metalloproteinases in atherosclerosis. 相似文献
2.
Silverthorn DU 《Advances in physiology education》2006,30(4):135-140
The Claude Bernard Distinguished Lectureship of the Teaching of Physiology Section is presented annually at the Experimental Biology meeting. The lectureship is named for Prof. Claude Bernard, the experimental physiologist who is credited with introducing the concept of homeostasis. The 2006 Claude Bernard Distinguished Lecture was given by Dr. Dee U. Silverthorn from the University of Texas at Austin, TX. 相似文献
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Joseph J. Tumilowicz 《In vitro cellular & developmental biology. Plant》1990,26(12):1144-1150
Summary Distinct, sequential events occurring during the destruction and simultaneous regrowth of human arterial smooth muscle cell
(SMC) cultures infected with cytomegalovirus (CMV, AD169 strain) were characterized. The events were influenced by the typical
phenotypic diversity reflecting relative states of differentiation of the SMC cultures. Progenitors of regeneration were a
surviving population of small, undifferentiated or relatively undifferentiated SMCs. As these cells reached confluence focally,
the number of cells reactive with antismooth muscle serum, i.e. differentiating, increased, and in some postconfluent foci
the organization of SMCs resembled the topography of uninfected cultures. Thus, infected SMC cultures had a limited capacity
to repopulate, to organize typically, and to differentiate. However, continuing cytopathic effects gradually destroyed much
of the regrowth, and a relatively large, nondividing SMC with prominent cytoplasmic filaments, similar to SMCs in terminal,
uninfected cultures, predominated. Infected cultures consisting overwhelmingly of the large terminal phenotype were far less
productive of infectious CMV than cultures populated by SMCs with continuing capacity to divide. Gradually, cultures consisting
of the terminal phenotype deteriorated as a result of sporadic cytopathic effects of CMV and an effect resembling “senescent”
degeneration in uninfected, nondividing cultures in late passage. The infected, terminal phenotype could be a latent or persistent
source of CMV antigen or nucleic acid-positive cells detected by different investigators in normal and in atheromatous, human
tissue, assuming that it exists and survives for an extended period in vivo after infection of vascular SMC.
The derivation of smooth muscle cell lines used in this investigation was supported through National Research and Demonstration
Center grant HL-17269-07 from the National Heart, Lung and Blood Institute, Bethesda, MD. 相似文献
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7.
It has recently been demonstrated that several neuropeptides can affect cell growth. The mammalian tachykinins substance P and neurokinin A, which are present in peripheral sensory neurons, stimulate growth of cultured connective tissue cells. Substance P-like immunoreactivity has been demonstrated in neuroblastoma cell lines. Neuroblastoma cells also produce other neuropeptides, among them vasoactive intestinal polypeptide (VIP). We report here that VIP is a potent inhibitor of serum-induced DNA synthesis in cultured smooth muscle cells (SMC), whereas no growth-inhibition was seen in SMC exposed to neurokinin A, calcitonin-gene related peptide, neuropeptide Y, somatostatin, or cholecystokinin. The growth-inhibitory effect of VIP was closely related to its ability to induce formation of cyclic AMP. Our results raise the possibility that peptides released by neurons, endocrine cells, as well as by transformed cells, may not only function as mitogens but also as inhibitory modulators of cell growth. 相似文献
8.
It has been known that benzimidazol-4,7-diones have antiproliferative activity against various cancer cell lines. Recently, we have also reported that these compounds strongly inhibited the proliferation of vascular smooth muscle cell (SMC) and human umbilical vein endothelial cells (HUVECs). Although benzimidazol-4,7-diones have important biological activities, the molecular mechanism of the compounds in these cells remains to be elucidated. In order to investigate the anti-proliferation mechanism of the compounds in smooth muscle cell, we selected 6-anilino-6-chloro-5-chloro-1H-benzo{d}midazole-4,7-dione (BUD-0203) among 12 benzimidazol-4,7-dione derivatives and examined its antiproliferative effects. Phosphorylation of the extracellular-signal regulated kinase (ERK) reached a maximal level at 1h after treatment with BUD-0203 and was sustained during the examined period. We also observed that phosphorylation of p38 reached a maximal level at 4h and decreased to control levels after 8h. These results showed that BUD-0203 sustainedly activated MAP kinase pathways in SMC. However, this compound did not induce cell cycle arrest in G1 or G2 phase in these cells. We also demonstrated that BUD-0203 not only induced apoptosis of SMC, but it also strongly inhibited SMC migration induced by platelet-derived growth factor (PDGF) or serum. Taken together, our experiments indicate that benzimidazol-4,7-diones induce apoptosis of smooth muscle cell via simultaneously prolonged activation of MAP kinase pathways including ERK, p38, and JNK/SAPK, similar with the apoptosis mechanism reported previously. 相似文献
9.
K. Shankar Narayan Neima Hendren 《In vitro cellular & developmental biology. Plant》1976,12(6):436-441
Summary Conditioned media of 21 mammalian cell lines have been examined by electron microscopic and biochemical techniques for the
presence of a minute ring-shaped nucleoprotein particle (RSP). Electron microscopy showed the presence of RSP in 20 of the
21 cell lines tested. Comparative analyses of six cell cultures for radio-active RSP, following3H-thymidine incorporation, suggested that quantitative differences exist in the amount of RSP detectable in conditioned media
of normal and pathologic cells. The cell lines tested included cells of different morphologies, origin and function.
Supported by a research grant from Claremont Foundation, Inc., Claremont, California. 相似文献
10.
Karen Zelinsky-Papez Timothy H. Carter Jay A. Zimmerman 《In vitro cellular & developmental biology. Plant》1987,23(2):118-122
Summary To evaluate the role of animal age in chemically induced transformation, pancreatic cells were grown in culture 6 to 8 wk
after injecting mice at either 6 or 22 mo. of age with a single dose ofN-methyl-N-nitrosourea (NMU). The cell type and the frequency with which lines were obtained from aged animals paralleled the frequency
and pattern of tumor induction by NMU in vivo. Outgrowth of pancreatic explants from young animals required the presence of
the tumor promoter 12-o-tetradecanoyl-phorbol-13-acetate to establish continuously growing cell lines. Whereas NMU alone produced
lines from aged mice, the promoter did not increase the frequency with which continuous lines were recovered from the aged
animals. Of eight cloned cell lines (four young and four old), all had characteristics of transformed mouse pancreatic acinar
cells when tested for lectin binding, lactate dehydrogenase isozyme pattern, chromosome number, and anchorage-independent
growth. Cell lines derived from aged animals were slower growing and had higher chromosome numbers than lines derived from
their younger counterparts. 相似文献
11.
《Cell cycle (Georgetown, Tex.)》2013,12(8):1048-1054
Paclitaxel (PTX), a microtubule-active drug, causes mitotic arrest leading to apoptosis in certain tumor cell lines. Here we investigated the effects of PTX on human arterial smooth muscle cell (SMC) cells. In SMC, PTX caused both (a) primary arrest in G1 and (b) post-mitotic arrest in G1. Post-mitotic cells were multinucleated (MN) with either 2C (near-diploid) or 4C (tetraploid) DNA content. At PTX concentrations above12 ng/ml, MN cells had 4C DNA content consistent with the lack of cytokinesis during abortive mitosis. Treatment with 6-12 ng/ml PTX yielded MN cells with 2C DNA content. Finally, 1-6 ng/ml of PTX, the lowest concentrations that affected cell proliferation, caused G1 arrest without multinucleation. It is important that PTX did not cause apoptosis in SMC. The absence of apoptosis could be explained by mitotic exit and G1 arrest as well as by low constitutive levels of caspase expression and by p53 and p21 induction. Thus, following transient mitotic arrest, SMC exit mitosis to form MN cells. These post-mitotic cells were subsequently arrested in G1 but maintained normal elongated morphology and were viable for at least 21 days. We conclude that in SMC PTX causes post-mitotic cell cycle arrest rather than cell death. 相似文献
12.
Lawrence E. DeBault Eduardo Henriquez Michael N. Hart Pasquale A. Cancilla 《In vitro cellular & developmental biology. Plant》1981,17(6):480-494
Summary An endothelial cell line has been established from a primary culture of cerebral microvessels isolated from Swiss-Webster
mice. The microvessels were isolated by a mechanical dispersion and filtration technique. The cells that emerged from these
microvessels, maintained in organoid cultures, proliferated and formed plaques of a single or mixed cell type. The endothelial
cell line, designated ME-2, was isolated from one such morphologically homogeneous cell plaque, using both cloning ring techniques
and C6 glioma-conditioned medium.
An endothelial specific antiserum was made in rabbits and was used immunocytochemically to confirm the cell type of origin
of the ME-2 cell line. Not only did the cell type specific antiserum react exclusively with endothelial cells in vivo, but
in the brain the antiserum localized preferentially to the luminal membrane of the endothelium.
The ME-2 endothelial cells have retained several of their unique properties such as cytomorphology, growth characteristics,
and cell type specific surface antigens throughout the life of the line (in one case 40 passages before senescence).
This work was supported in part by an Arteriosclerosis Specialized Center of Research grant from the National Heart, Lung
and Blood Institute, National Institutes of Health, Grant HL-14230, and Grant 584-127703 from the Veterans Adminsitration.
This paper is dedicated to the memory of Steve Frommes, Electron Microscopist and Photographer. 相似文献
13.
Intermediate filament expression and lifespan potential in human somatic cell hybrids 总被引:1,自引:0,他引:1
Clive L. Bunn Frank A. White W. Michael O’Guin Roger H. Sawyer Loren W. Knapp 《In vitro cellular & developmental biology. Plant》1985,21(12):716-720
Summary Limited lifespan human diploid fibroblast cells have been fused with the HeLa derived cell line HEB 7A which possesses transformed
growth characteristics and unlimited division potential. HEB 7A expresses keratin intermediate filaments, while the fibroblast
cells express only vimentin intermediate filaments. Independently arising clones of hybrids were examined for the presence
of keratin by indirect immunofluorescence. Of 11 limited lifespan hybrids, all were keratin negative and possessed the growth
characteristics of the fibroblast parent. Of 8 transformed hybrids, 6 arising early after fusion and 2 arising late, all were
keratin-positive and simultaneously expressed the transformed growth characteristics of loss of density dependent growth inhibition,
low serum dependence, and anchorage independence. It is concluded that the growth properties of these hybrids are associated
with the type of intermediate filament expressed. The intermediate filament expression is therefore a marker of proliferative
potential in these hybrids.
This work was supported by grant no. AG 02664 from NIA (to C.L.B.) and by grant nos. 1R01 HD 18129-01 from NIH and PCM83-09068
from NSF (to R.H.S.).
Editor’s Statement The tight correlation between the expression of the intermediate filaments of the immortal parent in hybrids
of limited lifespan fibroblasts and HeLa cells with the transformed phenotype is of interest. It may offer important clues
to the mechanism involved in cellular senescence. Gordon H. Sato 相似文献
14.
Simian adenovirus 7 gave an abortive infection in simian marmoset lymphoblastoid cells, B 95-8 and M 81 (transformed by Epstein Barr Virus) whereas non transformed simian lymphocytes could not replicate this virus. Electron dense incomplete particles with a lower density than standard virus in CsCl gradients were isolated. Virus yields were low and the percentage of cells containing viral antigen as measured by immunofluorescence was 0.01% for B 95-8 cells and still less for the M 81 cells. After a single passage in either lymphoblastic cell lines, they had a reduced oncogenicity in vivo. The polypeptide pattern analysis by PAGE showed some modifications. 相似文献
15.
Ehrlich HP Allison GM Page MJ Kolton WA Graham M 《Journal of cellular physiology》2000,182(2):303-309
Intestinal smooth muscle cells (SMC) produce the fibrotic tissue, strictures, that characterize Crohn's disease. These SMC change their phenotype from a contractile muscle form to an inflammation-responsive form that migrates and synthesizes a collagen matrix. It is postulated that the inflammatory responsive SMC form associates differently with its surrounding collagen matrix compared to the normal SMC form. SMC derived from Crohn's diseased and uninvolved bowel were sustained in cell culture. Cultured SMC incorporated in collagen lattices have the capacity to reduce the size of that lattice, referred to as lattice contraction. At day 2, Crohn's SMC-populated collagen lattices were reduced to 21% of their initial area, while non-Crohn's SMC collagen lattices were reduced to 8%. Crohn's SMC demonstrate retarded lattice contraction compared to non-Crohn's SMC. When grown in monolayer culture, Crohn's-derived SMC cover 30% more area than non-Crohn's SMC. By Western blot analysis Crohn's SMC express more gelsolin, an actin-binding protein found elevated in cells exhibiting increased cell motility. Was the increased expression of gelsolin related to retarded collagen lattice contraction? Intracellular levels of gelsolin were elevated by the electroporation of plasma gelsolin protein into suspended non-Crohn's SMC. When incorporated in collagen lattices, gelsolin loaded cells showed retarded lattice contraction compared to SMC loaded with albumin. Crohn's SMC show increased expression of gelsolin, which may be associated with a diminished capacity to reorganize collagen fiber bundles. It is suggested that increased concentrations of gelsolin in Crohn's SMC is consistent with enhanced cell migration as a consequence of the inflammatory state of Crohn's diseased intestine. 相似文献
16.
Yang B Gwozdz T Dutko-Gwozdz J Bolotina VM 《American journal of physiology. Cell physiology》2012,302(5):C748-C756
Store-operated Ca(2+) entry (SOCE) is important for multiple functions of vascular smooth muscle cells (SMC), which, depending of their phenotype, can resemble excitable and nonexcitable cells. Similar to nonexcitable cells, Orai1 was found to mediate Ca(2+)-selective (CRAC-like) current and SOCE in dedifferentiated cultured SMC and smooth muscle-derived cell lines. However, the role of Orai1 in cation-selective store-operated channels (cat-SOC), which are responsible for SOCE in primary SMC, remains unclear. Here we focus on primary SMC, and assess the role of Orai1 and Ca(2+)-independent phospholipase A(2) (iPLA(2)β, or PLA2G6) in activation of cat-SOC current (I(cat-SOC)), SOCE, and SMC proliferation. Using molecular, electrophysiological, imaging, and functional approaches, we demonstrate that molecular knockdown of either Orai1 or iPLA(2)β leads to similar inhibition of the whole cell cat-SOC current and SOCE in primary aortic SMC and results in significant reduction in DNA synthesis and impairment of SMC proliferation. This is the first demonstration that Orai1 and iPLA(2)β are equally important for cat-SOC, SOCE, and proliferation of primary aortic SMC. 相似文献
17.
Apoptosis repressor with a CARD domain (ARC), which has been shown to protect against oxidative stress-induced apoptosis, was initially found to be highly expressed in terminally differentiated tissues like heart and skeletal muscle. Recently, we and others have found that ARC is also expressed at high levels in multiple cancer tissues and cell lines. Here, we compared the regulation of ARC in response to oxidative stress between cancer cells and other types of cells. Similar to cardiomyocyte cell line H9c2 cells, cancer cells with reduced ARC expression were significantly more sensitive to oxidative stress. However, oxidative stress did not down-regulate ARC expression in cancer cells as it did in H9c2 cells. We further found that in H9c2 cells oxidative stress regulates ARC protein expression post-translationally through proteasome-mediated degradation. In cancer cell line HeLa, the majority of ARC exists in phosphorylated state in the absence of oxidative stress, whereas in H9c2 cells only marginal amount of ARC was phosphorylated under similar conditions. Our data suggest that the high level of ARC protein and the constitutive phosphorylation of ARC in cancer cells may play an important role in the protection of cancer cells against oxidative stress. 相似文献
18.
Aust G Wandel E Boltze C Sittig D Schütz A Horn LC Wobus M 《Cell and tissue research》2006,324(1):139-147
CD97, an epidermal growth factor (EGF)-TM7 receptor, is not restricted to hematopoetic and carcinoma cells but is also found
on smooth muscle cells (SMC). We have examined its location and biochemical structure in various normal and tumorigenic SMC-containing
tissues. SMC of the urinary bladder, lung bronchi and bronchioles, myometrium, and gastrointestinal tract were immunohistologically
stained by using monoclonal antibodies (mabs) to the CD97 stalk region (CD97stalk). Mabs directed against an N-glycosylation-dependent epitope within the EGF-domains (CD97EGF) did not bind to normal SMC. Vascular SMC, which was also CD97EGF-negative, showed further CD97 heterogeneity. Only a few, if any, SMC from the aorta or elastic arteries of the systemic circulation
were positive for CD97 mRNA and therefore also for CD97stalk. CD97stalk-positive SMC were slightly more numerous in muscular and peripheral arteries. In contrast, most venous SMC expressed CD97stalk. A comparison with other SMC molecules revealed a similar but not identical staining pattern for CD97stalk and desmin. Further CD97 heterogeneity was observed during SMC transformation. All leiomyomas (n=5) and nine out of 21 leiomyosarcomas were positive for both CD97stalk and CD97EGF. As expected, CD97EGF-positive SMC tumors expressed partly N-glycosylated CD97. Seven out of 21 leiomyosarcomas were completely devoid of CD97.
Thus, CD97 showed variable expression in vascular and biochemical modification in tumorigenic SMC, suggesting that the function
of the molecule is specific for the SMC subtype.
This study was supported by a joint grant from the German Research Council (DFG; project AU 132/3-1) and by the Interdisziplinary
Center of Clinical Research (IZKF) Leipzig at the Faculty of Medicine, University of Leipzig (project D6). E. Wandel is a
fellow of the IZKF. 相似文献
19.
Luechtenborg B Hofnagel O Weissen-Plenz G Severs NJ Robenek H 《European journal of cell biology》2008,87(2):91-99
Macrophages (MPhi) and smooth muscle cells (SMC) are transformed into foam cells by massive accumulation of modified lipoproteins during atherogenesis. It is known that class AI/II scavenger receptors participate in the foam cell formation of MPhi. The mechanism of lipid accumulation in SMC is however unknown. Therefore, we investigated if class AI/II scavenger receptors mediate the uptake of modified lipoproteins in SMC. Additionally, we examined the influence of MPhi and proinflammatory cytokines in this process. Our flow cytometric experiments revealed significant uptake of DiI-AcLDL in SMC. This uptake was markedly enhanced by IL-1alpha and TNF-alpha, whereas cocultured MPhi decreased the uptake of DiI-AcLDL in SMC. Competition and blocking experiments were performed to enlighten the role of class AI/II scavenger receptors. The competition experiments showed that surplus NatLDL, a ligand not known to interact with class AI/II scavenger receptors, caused a drastically decreased uptake of DiI-AcLDL in SMC. Additionally, blocking of class AI/II scavenger receptors with antibody 2F8 did not influence the uptake of DiI-AcLDL in SMC. Furthermore, fluorescence microscopic double staining of human coronary arteries with early, intermediate and advanced atherosclerotic lesions showed no colocalization of class AI scavenger receptors with SMC. These results indicate that class AI/II scavenger receptors play only a minor role in the uptake of modified lipoproteins in SMC. We suggest that SMC foam cell formation is mainly mediated by other receptors than class AI/II scavenger receptors. 相似文献
20.
Bernard Tandler 《Cell and tissue research》1965,68(6):852-863
Summary Myoepithelial cells in the human submaxillary gland are stellate in form, with long, tapering processes. They are interposed between the base of the secretory cells and the basement membrane, and are bound to the secretory cells by desmosomes. Their cytoplasm contains numerous myofilaments measuring approximately 40 Å in diameter, which frequently aggregate to form structures similar to the dark bodies seen in smooth muscle cells. The myofilaments are anchored to the plasma membrane by attachment devices. The myoepithelial cells are often accompanied by cells which have a similar shape, but possess an extremely electron-lucent cytoplasm that contains almost no organelles or inclusions. Electron microscopical observations indicate that these cells, termed clear cells, are transformed directly into myoepithelium, since all morphological intergrades between the two cell types have been recognized.This work was supported in part by a grant from the Henry Spenadel Trust. The technical assistance of Mr. Roy R. Keppie and Mrs. Mona Brandreth is gratefully acknowledged. 相似文献