Lockdown of mitochondrial Ca2+ extrusion and subsequent resveratrol treatment kill HeLa cells by Ca2+ overload

Abstract Background : The aberrant STAT3 signaling promotes initiation and progression of human cancers by either inhibiting apoptosis or inducing cell proliferation, angiogenesis, invasion, and metastasis. This study aims to investigate the role of STAT3 and its phosphorylation status in resveratrol-mediated suppression of cervical cancer. Methods : The effects of resveratrol on cervical tumor growth were also determined by examining the tumor tissues and their histological changes and the volume and weight of tumor tissues grown from Hela cells injected in female athymic BALB/C nude mice following the pretreatment regimen and the treatment regimen. Resveratrol structure and targets interaction virtual screening was performed using molecular docking program Autodock Vina. The phosphorylated STAT3, EMT molecular markers and ECM degradation enzymes protein levels were determined using Western blotting. Results : Proliferation and the colony formation of Hela cells were inhibited after resveratrol treatment in both dose- and time- dependent manner. Resveratrol inhibited migration and invasion of Hela cells. Molecular docking analysis showed that resveratrol interacted with STAT3 at a pocket composed of 11 amino acidic residues. Treatment with resveratrol resulted in decreases in the level of phosphorylation of STAT3 at Tyr705 but not Ser727, and no obvious changes in the protein level of STAT3 in Hela cells and SiHa cells. Pretreatment or treatment with resveratrol resulted in decreases in the IL-6-induced phosphorylation level of STAT3Tyr705 in Hela cells and SiHa cells, compared with those of treatment with IL-6. Reduced STAT3Tyr705 phosphorylation after STAT3 inhibitors S3I201 enhanced inhibition of invasion potential of Hela cells and SiHa cells treated with Resveratrol. Resveratrol decreases the N-Cadherin, Vimentin, MMP-3, MMP-13 protein level and increases the E-Cadherin protein level in a dose-dependent manner. The tumor size, volume, and weight, and the N-Cadherin, Vimentin, MMP-3, and MMP-13 protein level were significantly decreased, and the E-Cadherin protein level was increased, and the tumors were histologically damaged as revealed by H&E staining in both the resveratrol pretreatment group and the resveratrol treatment group and the magnitude of changes was higher in the former than that of the latter. Conclusion : Resveratrol inhibits growth and metastatic potential of cervical cancer through blocking STAT3Tyr705 phosphorylation.

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Platinum Compounds as Stains for Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100051165 ◽

1974 ◽

Vol 32 ◽

pp. 230-231

Author(s):

S. K. Aggarwal ◽

P. McAllister ◽

R. W. Wagner ◽

B. Rosenberg

Keyword(s):

Electron Microscopy ◽

Hela Cells ◽

Uranyl Acetate ◽

Sarcoma 180 ◽

Platinum Compounds ◽

Kb Cells ◽

En Bloc ◽

Human Lymphoma ◽

Ultrathin Sections ◽

Blue Coloration

Uranyl acetate has been used as an electron stain for en bloc staining as well as for staining ultrathin sections in conjunction with various lead stains (Fig. 1). Present studies reveal that various platinum compounds also show promise as electron stains. Certain platinum compounds have been shown to be effective anti-tumor agents. Of particular interest are the compounds with either uracil or thymine as one of the ligands (cis-Pt(II)-uracil; cis-Pt(II)-thymine). These compounds are amorphous, highly soluble in water and often exhibit an intense blue coloration. These compounds show enough electron density to be used as stains for electron microscopy. Most of the studies are based on various cell lines (human AV, cells, human lymphoma cells, KB cells, Sarcoma-180 ascites cells, chick fibroblasts and HeLa cells) while studies on tissue blocks are in progress.

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Electron Microscopy of Mycoplasma Pneumoniae

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100065341 ◽

1971 ◽

Vol 29 ◽

pp. 258-259 ◽

Cited By ~ 1

Author(s):

E. S. Boatman ◽

G. E. Kenny

Keyword(s):

Electron Microscopy ◽

Light Microscopy ◽

Growth Phase ◽

Hela Cells ◽

Sulfonic Acid ◽

Gamma Globulin ◽

Horse Serum ◽

Morphological Aspects ◽

The Family

Information concerning the morphology and replication of organism of the family Mycoplasmataceae remains, despite over 70 years of study, highly controversial. Due to their small size observations by light microscopy have not been rewarding. Furthermore, not only are these organisms extremely pleomorphic but their morphology also changes according to growth phase. This study deals with the morphological aspects of M. pneumoniae strain 3546 in relation to growth, interaction with HeLa cells and possible mechanisms of replication.The organisms were grown aerobically at 37°C in a soy peptone yeast dialysate medium supplemented with 12% gamma-globulin free horse serum. The medium was buffered at pH 7.3 with TES [N-tris (hyroxymethyl) methyl-2-aminoethane sulfonic acid] at 10mM concentration. The inoculum, an actively growing culture, was filtered through a 0.5 μm polycarbonate “nuclepore” filter to prevent transfer of all but the smallest aggregates. Growth was assessed at specific periods by colony counts and 800 ml samples of organisms were fixed in situ with 2.5% glutaraldehyde for 3 hrs. at 4°C. Washed cells for sectioning were post-fixed in 0.8% OSO4 in veronal-acetate buffer pH 6.1 for 1 hr. at 21°C. HeLa cells were infected with a filtered inoculum of M. pneumoniae and incubated for 9 days in Leighton tubes with coverslips. The cells were then removed and processed for electron microscopy.

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Use of Uranium-Labeled Antibodies for Electron Microscopic Localization of Various Antigens in Mammalian Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100060696 ◽

1967 ◽

Vol 25 ◽

pp. 136-137

Author(s):

J. P. Petrali ◽

E. J. Donati ◽

L. A. Sternberger

Keyword(s):

Endoplasmic Reticulum ◽

Hela Cells ◽

Mammalian Cells ◽

Specific Antiserum ◽

Electron Microscopic ◽

E Coli ◽

Cytoplasmic Membranes ◽

Viral Progeny ◽

Ultrathin Sections ◽

Electron Microscopic Localization

Specific contrast is conferred to subcellular antigen by applying purified antibodies, exhaustively labeled with uranium under immunospecific protection, to ultrathin sections. Use of Seligman’s principle of bridging osmium to metal via thiocarbohydrazide (TCH) intensifies specific contrast. Ultrathin sections of osmium-fixed materials were stained on the grid by application of 1) thiosemicarbazide (TSC), 2) unlabeled specific antiserum, 3) uranium-labeled anti-antibody and 4) TCH followed by reosmication. Antigens to be localized consisted of vaccinia antigen in infected HeLa cells, lysozyme in monocytes of patients with monocytic or monomyelocytic leukemia, and fibrinogen in the platelets of these leukemic patients. Control sections were stained with non-specific antiserum (E. coli).In the vaccinia-HeLa system, antigen was localized from 1 to 3 hours following infection, and was confined to degrading virus, the inner walls of numerous organelles, and other structures in cytoplasmic foci. Surrounding architecture and cellular mitochondria were unstained. 8 to 14 hours after infection, antigen was localized on the outer walls of the viral progeny, on cytoplasmic membranes, and free in the cytoplasm. Staining of endoplasmic reticulum was intense and focal early, and weak and diffuse late in infection.

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Exogeneous factors are not necessary in attachment, spreading and polarization of hela cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100082418 ◽

1978 ◽

Vol 36 (2) ◽

pp. 310-311

Author(s):

W. Liebrich

Keyword(s):

Hela Cells ◽

Calf Serum ◽

Glass Tube ◽

Serum Free Medium ◽

Nacl Solution ◽

Free Medium ◽

Minimum Essential Medium ◽

Serum Free ◽

All Solutions ◽

Glass Support

HeLa cells were grown for 2-3 days in EAGLE'S minimum essential medium with 10% calf serum (S-MEM; Seromed, München) and then incubated for 24 hours in serum free medium (MEM). After detaching the cells with a solution of 0. 14 % EDTA and 0. 07 % trypsin (Difco, 1 : 250) they were suspended in various solutions (S-MEM = control, MEM, buffered salt solutions with or without Me++ions, 0. 9 % NaCl solution) and allowed to settle on glass tube slips (Leighton-tubes). After 5, 10, 15, 20, 25, 30, 1 45, 60 minutes 2, 3, 4, 5 hours cells were prepared for scanning electron microscopy as described by Paweletz and Schroeter. The preparations were examined in a Jeol SEM (JSM-U3) at 25 KV without tilting.The suspended spherical HeLa cells are able to adhere to the glass support in all solutions. The rate of attachment, however, is faster in solutions without serum than in the control. The latter is in agreement with the findings of other authors.

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Human Adenovirus Uptake by Preirnplantation Mouse Embryos

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094607 ◽

1972 ◽

Vol 30 ◽

pp. 268-269

Author(s):

D. G. Chase ◽

W. Winters ◽

L. Piko

Keyword(s):

Hela Cells ◽

Human Adenovirus ◽

Mouse Embryos ◽

Equilibrium Density ◽

Cell Stage ◽

Virus Attachment ◽

Preimplantation Mouse Embryos ◽

Embryo Culture Medium ◽

Preimplantation Mouse ◽

Mouse Cells

Although the outlines of human adenovirus entry and uncoating in HeLa cells has been clarified in recent electron microscope studies, several details remain unclear or controversial. Furthermore, morphological features of early interactions of human adenovirus with non-permissive mouse cells have not been extensively documented. In the course of studies on the effects of human adenoviruses type 5 (AD-5) and type 12 on cultured preimplantation mouse embryos we have examined virus attachment, entry and uncoating. Here we present the ultrastructural findings for AD-5.AD-5 was grown in HeLa cells and purified by successive velocity gradient and equilibrium density gradient centrifugations in CsCl. After dialysis against PBS, virus was sedimented and resuspended in embryo culture medium. Embryos were placed in culture at the 2-cell stage in Brinster's medium.

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Temporal and spatial interrelationships of confronting cisternae to nuclear envelope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100125051 ◽

1992 ◽

Vol 50 (1) ◽

pp. 930-931

Author(s):

John R. Palisano

Keyword(s):

Endoplasmic Reticulum ◽

Nuclear Envelope ◽

Tumor Cells ◽

Hela Cells ◽

Microscopic Investigation ◽

Electron Microscopic Investigation ◽

Serial Sections ◽

Electron Microscopic ◽

Fetal Rat ◽

Temporal And Spatial

Although confronting cistemae (CC) have been observed in a variety of tumor cells and normal fetal rat, mouse, and human epithelial tissues, little is known about their origin or role in mitotic cells. While several investigators have suggested that CC arise from nuclear envelope (NE) folding back on itself during prophase, others have suggested that CC arise when fragments of NE pair with endoplasmic reticulum. An electron microscopic investigation of 0.25 um thick serial sections was undertaken to examine the origin of CC in HeLa cells.

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