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In Vitro Investigations on the Toxicity and Cell Death Induced by Tamoxifen on Two Non-Breast Cancer Cell Types.

Majumdar SK, Valdellon JA, Brown KA - J. Biomed. Biotechnol. (2001)

Bottom Line: Tamoxifen treatment demonstrated an inhibitory effect on HeLa cell multiplication at lower concentrations and toxicity at higher concentrations and longer treatment durations.Surface ultrastructure of tamoxifen treated cells examined under the SEM revealed abnormalities such as membrane blebbing, holes, and cytoplasmic extrusions, all of which are characteristics of programmed cell death (apoptosis).Redistribution of the membrane phospholipid phosphatidylserine (PS) from the protoplasmic surface of the plasma membrane to the cell surface was identified using annexin V-enhanced green fluorescent protein (EGFP) in tamoxifen treated MEL BB-88 cells, a general feature of cells undergoing apoptosis.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
Tamoxifen, a potent anticancer agent known to interrupt the enhanced estrogen activity of malignant mammary gland cells, was recently approved by the Food and Drug Administration (FDA)for the treatment of breast cancer. In this investigation, the toxic effects of tamoxifen were evaluated through cell multiplication, and cytological, surface ultrastructural, and biochemical studies on human cervical carcinoma cells (HeLa)and/or murine erythroleukemic (MEL) cells (BB-88). Tamoxifen treatment demonstrated an inhibitory effect on HeLa cell multiplication at lower concentrations and toxicity at higher concentrations and longer treatment durations. The drug also triggered morphological and biochemical changes as revealed by light microscopy, scanning electron microscopy (SEM), fluorescence microscopy, Nucleosome ELISA, and the DNA smear pattern. Cytological observations showed nuclear condensation, cell shrinkage, multinucleation, and apoptotic bodies. Surface ultrastructure of tamoxifen treated cells examined under the SEM revealed abnormalities such as membrane blebbing, holes, and cytoplasmic extrusions, all of which are characteristics of programmed cell death (apoptosis). Redistribution of the membrane phospholipid phosphatidylserine (PS) from the protoplasmic surface of the plasma membrane to the cell surface was identified using annexin V-enhanced green fluorescent protein (EGFP) in tamoxifen treated MEL BB-88 cells, a general feature of cells undergoing apoptosis. Tamoxifen treated cells demonstrated internucleosomal damages of the genomic DNA and DNA fragmentations, evidenced by an increase in free nucleosomes, and distinctive DNA smear patterns on the agarose gel.

No MeSH data available.


Related in: MedlinePlus

Immunofluorascence staining with annexin V-enhanced green fluorescent protein (EGFP) and propidium iodide (PI). (a), (b) Control BB-88 cells (24 hours) showing red staining due to PI throughout the cell (× 400). (c), (d) Tamoxifen treated BB-88 cells (8 hours, 5 μg/ml) showing annexin V-EGFP green staining in the plasma membrane ((c) × 400, (d) × 900) and blebs on cells undergoing apoptosis.
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Figure 5: Immunofluorascence staining with annexin V-enhanced green fluorescent protein (EGFP) and propidium iodide (PI). (a), (b) Control BB-88 cells (24 hours) showing red staining due to PI throughout the cell (× 400). (c), (d) Tamoxifen treated BB-88 cells (8 hours, 5 μg/ml) showing annexin V-EGFP green staining in the plasma membrane ((c) × 400, (d) × 900) and blebs on cells undergoing apoptosis.

Mentions: Untreated BB-88 cells were spherical in shape and both the nucleus and the plasma membrane appeared red due to the nuclear counterstain propidium iodide (PI) (see Figures 5a and 5b). On occasions, dying or dead cells were detected in the untreated group with blebs and bound annexin V-enhanced fluorescent green protein (EGFP) which stained the cell membrane green. A higher proportion of tamoxifen treated BB-88 cells showed green staining in the plasma membrane resulted from the bound annexin- V protein to the redistributed phosatidylserine (PS) translocated on the surface of cells possibly undergoing apoptosis (see Figures 5c and 5d). Apoptosis is indicated by a green staining (EGFP) plasma membrane surrounding the lightly stained (PI) nucleus, as seen in Figures 5c and 5d [21].


In Vitro Investigations on the Toxicity and Cell Death Induced by Tamoxifen on Two Non-Breast Cancer Cell Types.

Majumdar SK, Valdellon JA, Brown KA - J. Biomed. Biotechnol. (2001)

Immunofluorascence staining with annexin V-enhanced green fluorescent protein (EGFP) and propidium iodide (PI). (a), (b) Control BB-88 cells (24 hours) showing red staining due to PI throughout the cell (× 400). (c), (d) Tamoxifen treated BB-88 cells (8 hours, 5 μg/ml) showing annexin V-EGFP green staining in the plasma membrane ((c) × 400, (d) × 900) and blebs on cells undergoing apoptosis.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC129061&req=5

Figure 5: Immunofluorascence staining with annexin V-enhanced green fluorescent protein (EGFP) and propidium iodide (PI). (a), (b) Control BB-88 cells (24 hours) showing red staining due to PI throughout the cell (× 400). (c), (d) Tamoxifen treated BB-88 cells (8 hours, 5 μg/ml) showing annexin V-EGFP green staining in the plasma membrane ((c) × 400, (d) × 900) and blebs on cells undergoing apoptosis.
Mentions: Untreated BB-88 cells were spherical in shape and both the nucleus and the plasma membrane appeared red due to the nuclear counterstain propidium iodide (PI) (see Figures 5a and 5b). On occasions, dying or dead cells were detected in the untreated group with blebs and bound annexin V-enhanced fluorescent green protein (EGFP) which stained the cell membrane green. A higher proportion of tamoxifen treated BB-88 cells showed green staining in the plasma membrane resulted from the bound annexin- V protein to the redistributed phosatidylserine (PS) translocated on the surface of cells possibly undergoing apoptosis (see Figures 5c and 5d). Apoptosis is indicated by a green staining (EGFP) plasma membrane surrounding the lightly stained (PI) nucleus, as seen in Figures 5c and 5d [21].

Bottom Line: Tamoxifen treatment demonstrated an inhibitory effect on HeLa cell multiplication at lower concentrations and toxicity at higher concentrations and longer treatment durations.Surface ultrastructure of tamoxifen treated cells examined under the SEM revealed abnormalities such as membrane blebbing, holes, and cytoplasmic extrusions, all of which are characteristics of programmed cell death (apoptosis).Redistribution of the membrane phospholipid phosphatidylserine (PS) from the protoplasmic surface of the plasma membrane to the cell surface was identified using annexin V-enhanced green fluorescent protein (EGFP) in tamoxifen treated MEL BB-88 cells, a general feature of cells undergoing apoptosis.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
Tamoxifen, a potent anticancer agent known to interrupt the enhanced estrogen activity of malignant mammary gland cells, was recently approved by the Food and Drug Administration (FDA)for the treatment of breast cancer. In this investigation, the toxic effects of tamoxifen were evaluated through cell multiplication, and cytological, surface ultrastructural, and biochemical studies on human cervical carcinoma cells (HeLa)and/or murine erythroleukemic (MEL) cells (BB-88). Tamoxifen treatment demonstrated an inhibitory effect on HeLa cell multiplication at lower concentrations and toxicity at higher concentrations and longer treatment durations. The drug also triggered morphological and biochemical changes as revealed by light microscopy, scanning electron microscopy (SEM), fluorescence microscopy, Nucleosome ELISA, and the DNA smear pattern. Cytological observations showed nuclear condensation, cell shrinkage, multinucleation, and apoptotic bodies. Surface ultrastructure of tamoxifen treated cells examined under the SEM revealed abnormalities such as membrane blebbing, holes, and cytoplasmic extrusions, all of which are characteristics of programmed cell death (apoptosis). Redistribution of the membrane phospholipid phosphatidylserine (PS) from the protoplasmic surface of the plasma membrane to the cell surface was identified using annexin V-enhanced green fluorescent protein (EGFP) in tamoxifen treated MEL BB-88 cells, a general feature of cells undergoing apoptosis. Tamoxifen treated cells demonstrated internucleosomal damages of the genomic DNA and DNA fragmentations, evidenced by an increase in free nucleosomes, and distinctive DNA smear patterns on the agarose gel.

No MeSH data available.


Related in: MedlinePlus