DNA fluorometric assay in 96-well tissue culture plates using Hoechst 33258 after cell lysis by freezing in distilled water

Rage, Randall; Mitchen, Jacque; Wilding, G.

doi:10.1016/0003-2697(90)90382-j

Cited by 378 publications

(228 citation statements)

References 3 publications

Supporting

Mentioning

223

Contrasting

Unclassified

Order By: Relevance

“…The number of cells was determined daily over a 5-day period using the Hoechst 33258 assay (Rago et al, 1990) with a GENios Pro plate reader (TECAN, Mannedorf, Switzerland) at the excitation wavelength of 360/35 nm and detection wavelength of 460/10 nm.…”

Section: Proliferation Assaymentioning

confidence: 99%

Cellular and molecular phenotypes of proliferating stromal cells from human carcinomas

et al. 2010

View full text Add to dashboard Cite

BACKGROUND: Stromal cells are a functionally important component of human carcinomas. The aim of this study was to obtain and characterise primary cultures of stromal cells from human carcinomas and the corresponding surrounding normal tissue. METHODS: Primary stromal cell cultures from tumours of lung, oesophagus and pancreas were obtained using a mild tissue dissociation method and a medium for culturing mesenchymal cells. Immunofluorescence staining and western blotting were used to analyse the expression of differentiation markers and selected known oncoproteins in the cell cultures obtained. RESULTS: A panel of stromal primary cultures was prepared from different human tumours and from matched normal cancer-free tissues. The in vitro proliferative potential of tumour-associated fibroblasts was shown to be higher than that of matched normal stromal cells. A mutational analysis of the TP53 and KRAS2 genes in a number of stromal cultures did not reveal known mutations in most cells of the cultures studied. Western blot analysis showed that stromal cells of lung tumours were characterised by a statistically significantly lower expression level of the p16 protein as compared with that in normal lung stromal cells. An important finding of our study was that, according to immunofluorescence assay, a fraction of fibroblast-like vimentin-positive cells in some tumour and normal stromal cell cultures expressed an epithelial marker -cytokeratins. CONCLUSION: Proliferating stromal cells from the carcinomas studied proved to be genetically normal cells with altered expression profiles of some genes involved in carcinogenesis, as compared with normal stromal cells. Epithelial-mesenchymal transition may lead to the emergence of transdifferentiated fibroblast-like cells in tumour stroma and in the tumour-surrounding tissue.

show abstract

Section: Proliferation Assaymentioning

confidence: 99%

Cellular and molecular phenotypes of proliferating stromal cells from human carcinomas

et al. 2010

View full text Add to dashboard Cite

show abstract

“…Cell number was analyzed by measuring DNA content with the fluorescent dye Hoechst 33258 (Sigma, St. Louis, MO) as described previously [28].…”

Section: Cell Proliferation Assaymentioning

confidence: 99%

The liver X receptor agonist T0901317 acts as androgen receptor antagonist in human prostate cancer cells

Chuu

Chen

Hiipakka

et al. 2007

Biochemical and Biophysical Research Communications

View full text Add to dashboard Cite

T0901317 is a potent non-steroidal synthetic liver X receptor (LXR) agonist. T0901317 blocked androgenic stimulation of the proliferation of androgen-dependent LNCaP 104-S cells and androgenic suppression of the proliferation of androgen-independent LNCaP 104-R2 cells, inhibited the transcriptional activation of an androgen-dependent reporter gene by androgen, and suppressed gene and protein expression of prostate specific antigen (PSA), a target gene of androgen receptor (AR) without affecting gene and protein expression of AR. T0901317 also inhibited binding of a radiolabeled androgen to AR, but inhibition was much weaker compared to the effect of the antiandrogens, bicalutamide and hydroxyflutamide. The LXR agonist T0901317, therefore, acts as an antiandrogen in human prostate cancer cells.

show abstract

“…PT was added to the medium for the indicated times at various concentrations. Cell number was estimated by a DNA fluorometric assay using the fluorochrome Hoechst 33258 (Rago et al, 1990).…”

Section: Hepatoma Cell Culture and Growth Inhibitionmentioning

confidence: 99%

Growth inhibitory actions of prothrombin on normal hepatocytes: Influence of matrix

Carr

Kar

Wang

et al. 2007

Cell Biology International

View full text Add to dashboard Cite

Most hepatomas have a defect in prothrombin carboxylation, and can secrete under-carboxylated prothrombin or des-γ-carboxy-prothrombin (DCP), the function of which is unknown. We considered that prothrombin-DCP axis might also be involved in growth control. Hepatocytes and hepatoma cells were treated with prothrombin, and DNA synthesis and cytoskeleton were studied. Prothrombin inhibited DNA synthesis in hepatocytes on fibronectin, but not collagen matrix. Hepatoma cell lines were not inhibited. We found that hepatoma cell matrix conferred resistance to hepatocytes. Prothrombin decreased fibronectin but not collagen amounts, but only in the presence of hepatocytes and not hepatoma cells, indicating that it has a differential action on matrix proteins. It also caused changes in cell shape and actin depolymerization. In vivo, there was a decrease in plasma prothrombin activity after a partial hepatectomy (PH) concomitant with a peak of DNA synthesis by the hepatocyte at 24 h after PH. Injection of warfarin at the time of PH, further inhibited PT activity and enhanced this 24 h peak of DNA synthesis. Furthermore, repeated injection of prothrombin lowered the peak DNA synthesis after PH. The data support the hypothesis that prothrombin can act as a hepatocyte growth inhibitor, likely at the level of fibronectin loss and result in cytoskeletal changes. Hepatomas resist this action, possibly due to their different matrix proteins. This represents a novel mechanism for growth regulation and provides a possible biological significance for the tumor marker DCP.

show abstract

DNA fluorometric assay in 96-well tissue culture plates using Hoechst 33258 after cell lysis by freezing in distilled water

Cited by 378 publications

References 3 publications

Cellular and molecular phenotypes of proliferating stromal cells from human carcinomas

Cellular and molecular phenotypes of proliferating stromal cells from human carcinomas

The liver X receptor agonist T0901317 acts as androgen receptor antagonist in human prostate cancer cells

Growth inhibitory actions of prothrombin on normal hepatocytes: Influence of matrix

Contact Info

Product

Resources

About