OBTAINING OF RADIORESISTANT VARIANTS OF HeLa AND DU145 CELL LINES

Papaverine (PPV) is an alkaloid isolated from the Papaver somniferum. Research has shown that PPV inhibits proliferation. However, several questions remain regarding the effects of PPV in tumorigenic cells. In this study, the influence of PPV was investigated on the proliferation (spectrophotometry), morphology (light microscopy), oxidative stress (fluorescent microscopy), and cell cycle progression (flow cytometry) in MDA-MB-231, A549, and DU145 cell lines. Exposure to 150 μM PPV resulted in time- and dose-dependent antiproliferative activity with reduced cell growth to 56%, 53%, and 64% in the MDA-MB-231, A549, and DU145 cell lines, respectively. Light microscopy revealed that PPV exposure increased cellular protrusions in MDA-MB-231 and A549 cells to 34% and 23%. Hydrogen peroxide production increased to 1.04-, 1.02-, and 1.44-fold in PPV-treated MDA-MB-231, A549, and DU145 cells, respectively, compared to cells propagated in growth medium. Furthermore, exposure to PPV resulted in an increase of cells in the sub-G1 phase by 46% and endoreduplication by 10% compared to cells propagated in growth medium that presented with 2.8% cells in the sub-G1 phase and less than 1% in endoreduplication. The results of this study contribute to understanding of effects of PPV on cancer cell lines.

Download Full-text

Diagnosis of human prostate carcinoma cancer stem cells enriched from DU145 cell lines changes with microscopic texture analysis in radiation and hyperthermia treatment using run-length matrix

International Journal of Radiation Biology ◽

10.1080/09553002.2017.1359429 ◽

2017 ◽

Vol 93 (11) ◽

pp. 1248-1256 ◽

Cited By ~ 1

Author(s):

Ali Abbasian Ardakani ◽

Jila Rajaee ◽

Samideh Khoei

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

Texture Analysis ◽

Cell Lines ◽

Prostate Carcinoma ◽

Du145 Cell ◽

Human Prostate ◽

Hyperthermia Treatment ◽

Run Length ◽

Human Prostate Carcinoma

Download Full-text

Deleted In Cancer 1 (DICE1): Search for a function in prostate carcinoma (PCa)

Journal of Clinical Oncology ◽

10.1200/jco.2007.25.18_suppl.15634 ◽

2007 ◽

Vol 25 (18_suppl) ◽

pp. 15634-15634

Author(s):

S. Filleur ◽

I. Wieland ◽

T. Nelius ◽

R. Kennedy ◽

W. De Riese ◽

...

Keyword(s):

Tumor Suppressor ◽

Mrna Expression ◽

Cell Lines ◽

Ectopic Expression ◽

Du145 Cell ◽

Prostate Tissue ◽

Normal Prostate ◽

Mammalian Expression ◽

Clonogenic Growth ◽

Normal Prostate Tissue

15634 Background: Recently, DICE1 gene (MIM 604331 ) was identified to colocalize with the microsatellite marker D13S284 in 13q14.3, a region frequently affected by allelic deletion in PCa. We previously showed that DICE1 mRNA expression is down-regulated in PCa cell lines compared to normal prostate tissue, due to DICE1 promoter hypermethylation; suggesting that DICE1 is a tumor suppressor gene. Methods: Human PCa cell lines PC3 and DU145 were lipo-transfected with mammalian expression plasmids encoding for the full length DICE1 gene or for its N- (APAI construct) and C-terminal (DEAD construct) fragments. The constructs expression was determined by semi-quantitative PCR and quantified by Real-time PCR. Clonogenic formation and apoptotic assays were performed. Results: The PCR analysis showed that the expression of DICE1, APAI and DEAD domains in transfected PC3 and DU145 cell lines was increased 30.6, 75.2 and 27.9 fold in PC3 cells and 4.3, 5 and 2.5 fold in DU145 cells, respectively, compared to non-transfected cells. The function analysis showed that the ectopic expression of DICE1 suppressed clonogenic growth of PC3 and DU145 cell lines. Surprisingly, we showed that like DICE1, DEAD and APAI inhibit the PC3 and DU145 clonogenic growth suggesting that both regions are involved in prostate tumor growth inhibition. The apoptosis assay could not show any DNA fragmentation activity for DICE1. Conclusions: We demonstrated that DICE1 is a tumor suppressor in PCa. DICE1 mRNA expression is down-regulated in PCa cell lines compared to normal prostate tissue and its ectopic expression in PCa cell lines inhibits their capacity to form clonogenic colonies in vitro. The functional analysis could not reveal any role of DICE1 in PCa apoptosis, suggesting that other molecular mechanisms are involved. No significant financial relationships to disclose.

Download Full-text

(EB) Virus: Latency and Expression in Transplanted and Cultured Human Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100065882 ◽

1971 ◽

Vol 29 ◽

pp. 368-369

Author(s):

B. G. Uzman ◽

M. M. Kasac ◽

H. Saito ◽

A. Krishan

Keyword(s):

Cell Lines ◽

Primary Cultures ◽

Virus Particles ◽

Barr Virus ◽

Virus Latency ◽

Virus Surveillance ◽

Cultured Human Cells ◽

Epstein Barr ◽

Serial Transplantation ◽

Tubular Arrays

In conjunction with the cultivation and transplantation of cells from human tumors by the Programs of Microbiology and Immunogenetics, virus surveillance by electron microscopy has been routinely employed. Of particular interest in this regard have been 3 cell lines cultured from lymph nodes or spleen of 2 patients with Hodgkin's disease and 1 patient with Letterer-Siwe's disease. Each of these cell lines when transplanted in Syrian hamster neonates conditioned with anti-lymphocyte serum grew as serially transplantable tumors; from such transplants of the 3 cell lines cell cultures were retrieved.Herpes type virus particles (Figs. 1, 2, 3) were found in the primary cultures of all three lines, in frozen thawed aliquots of same, and in cultures retrieved from their tumors growing by serial transplantation in hamsters. No virus was detected in sections of 25 of the serially transplanted tumors. However, in 10 such tumors there were repeated instances of tubular arrays in the cisternae of the endoplasmic reticulum (Fig. 4). On serologic examination the herpes virus was shown to be the Epstein-Barr virus.

Download Full-text

Comparison of Choriocarcinoma and Normal Placenta

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100065663 ◽

1971 ◽

Vol 29 ◽

pp. 324-325

Author(s):

John C. Garancis ◽

Roland A. Pattillo ◽

Robert O. Hussa ◽

Jon V. Straumfjord

Keyword(s):

Cell Lines ◽

Small Cells ◽

Tissue Cultures ◽

Glycogen Depletion ◽

Cell Surfaces ◽

Intercellular Spaces ◽

Concomitant Increase ◽

Gestational Choriocarcinoma ◽

Cytoplasmic Glycogen ◽

Normal Placenta

Two different cell lines (Be-Wo and Jar) of human gestational choriocarcinoma have been maintained in continuous tissue culture for a period of four and two years respectively without losing the ability to elaborate human chorionic gonadotropin (HCG). Tissue cultures, as revealed by electron microscopy, consisted of small cells with single nuclei. In some instances cell surfaces were provided with microvilli but more often the intercellular spaces were narrow and bridged by desmosomes. However, syncytium was not formed. Endoplasmic reticulum (ER) was poorly developed in both cell lines, except in some Be-Wo cells it was prominent. Golgi complex, lysosomes and numerous free ribosomes, as well as excessive cytoplasmic glycogen, were present in all cells (Fig. 1). Glycogen depletion and concomitant increase of ER were observed in many cells following a single dose of 10 ugm/ml of adrenalin added to medium (Fig. 2).

Download Full-text

Immunocytochemical localization of β1-4 galactosyltransferase in endometrial carcinoma cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162284 ◽

1990 ◽

Vol 48 (3) ◽

pp. 944-945

Author(s):

Ichiro Yamamoto ◽

Toshiaki Tachibana ◽

Hiroko Maruyama ◽

Noriyuki Komatsu ◽

Hiroyuki Kuramoto ◽

...

Keyword(s):

Endometrial Carcinoma ◽

Cell Lines ◽

Endometrial Adenocarcinoma ◽

Protein A ◽

Rabbit Antiserum ◽

Carcinoma Cells ◽

Affinity Column ◽

Antibody Technique ◽

Galactosyl Transferase ◽

C Terminus

We have paid attention to the alteration of glycosyltransferase in carcinoma cells, because it might be related to the malignancy of the cells. In this connection, localization of β1-4 galactosyl transferase (β1-4 Gal T) in human endometrial carcinoma cells was examined immunocytochemically using two kinds of cell lines, each of which showed different degree of differentiation.An antibody was purified from the rabbit antiserum against the synthetic peptide, IFNRLVFRGMSC (W89) of human β1-4 Gal T coupled with KLH (keyhole limpet hemocyanine) by protein A column and peptide-affinity column chromatography. The anti-W89 serum reacts to the C-terminus of human β 1-4 Gal T and to both membrane-bound and soluble forms of the enzyme. Cell line of well differentiated endometrial adenocarcinoma (I) and that of poorly differentiated endometrial adenocarcinoma (50B) were cultivated respectively in MEM medium containing 15% FCS and 2 mM glutamine for 4 d at 37°C under 5% CO2. The cells were fixed in a mixture of 4% paraformaldehyde and 0.1% glutaraldehyde in 0.1 M Soerensen’s phosphate buffer (pH 7.4) at 4°C for 30 min, washed with PBS, then freezed and thawed. The indirect method of the peroxidase- labeled antibody technique was used for immunocytochemistry of both LM and TEM on the cell lines. The cells were dehydrated in ethanol and embedded in TAAB 812. Ultrathin sections were observed under a TEM, JEM-100S.

Download Full-text

Absence of temporal ordering of apoptotic features in heat-shock treated leukemia and lymphoma cell lines

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100166257 ◽

1996 ◽

Vol 54 ◽

pp. 758-759

Author(s):

D. W. Fairbain ◽

M.D. Standing ◽

K.L. O'Neill

Keyword(s):

Heat Shock ◽

Cell Lines ◽

Chromatin Condensation ◽

Membrane Integrity ◽

Resistant Cell ◽

Membrane Blebbing ◽

Late Loss ◽

Induced Apoptosis ◽

Dying Cells ◽

In Cells

Apoptosis is a genetically defined response to physiological stimuli that results in cellular suicide. Features common to apoptotic cells include chromatin condensation, oligonucleosomal DNA fragmentation, membrane blebbing, nuclear destruction, and late loss of ability to exclude vital dyes. These characteristics contrast markedly from pathological necrosis, in which membrane integrity loss is demonstrated early, and other features of apoptosis, which allow a non-inflammatory removal of dead and dying cells, are absent. Using heat shock-induced apoptosis as a model for examining stress response in cells, we undertook to categorize a variety of human leukemias and lymphomas with regard to their response to heat shock. We were also interested in determining whether a common temporal order was followed in cells dying by apoptosis. In addition, based on our previous results, we investigated whether increasing heat load resulted in increased apoptosis, with particular interest in relatively resistant cell lines, or whether the mode of death changed from apoptosis to necrosis.

Download Full-text