An Assessment of the Immunofluorescence Technique as a Method for Demonstrating the Histological Localization of Tetrahydrocannabinol in Mammalian Tissues

Morley, Michael E.; Gee, David

doi:10.1520/jfs12200j

Cited by 5 publications

(4 citation statements)

References 10 publications

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“…Western blotting was performed as described previously . Total protein was prepared using extraction buffer comprising NaCl/P i containing 0.5% Triton X–100, 1 m m EDTA, 1 m m phenylmethyl sulfonyl fluoride, and complete protease inhibitors (Roche).…”

Section: Methodsmentioning

confidence: 99%

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Estrogen receptor α mediates proliferation of breast cancer MCF–7 cells via a p21/PCNA/E2F1‐dependent pathway

et al. 2014

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High expression of estrogen receptor a (ERa) is associated with a poor prognosis that correlates closely with cellular proliferation in breast cancer. However, the exact molecular mechanism by which ERa controls breast cancer cell proliferation is not clear. Here we report that ERa regulates the cell cycle by suppressing p53/p21 and up-regulating proliferating cell nuclear antigen (PCNA) and proliferation-related Ki-67 antigen (Ki-67) to promote proliferation of MCF-7 cells. In addition, 17-b-estradiol (E2) enhances ERa-induced proliferation of MCF-7 cells by stimulating expression of PCNA and Ki-67. Knockdown of ERa significantly affects PCNA/ Ki-67 and p53/p21 expression. Furthermore, ERa inhibits the transcriptional activity of p53/p21 in an estrogen response element-dependent manner. More importantly, we provide new evidence that ERa mediates proliferation of MCF-7 cells by up-regulating miR-17 to silence the expression of p21. Thus, these data provide new insights into the underlying effect of ERa on breast cancer proliferation.

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Section: Methodsmentioning

confidence: 99%

“…Immunocytochemistry assays were performed as described previously . After transfection with plasmids, the cells were fixed in 4% paraformaldehyde for 15 min, and then blocked with goat serum blocking solution for 20 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Estrogen receptor α mediates proliferation of breast cancer MCF–7 cells via a p21/PCNA/E2F1‐dependent pathway

et al. 2014

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“…Immunocytochemistry assays were performed as described previously (22). The cells after treatment were fixed in 4% paraformaldehyde for 15 min and then blocked with normal goat serum for 20 min at room temperature.…”

Section: Immunofluorescence Stainingmentioning

confidence: 99%

Human chorionic gonadotropin decreases human breast cancer cell proliferation and promotes differentiation

Liao

Wang

et al. 2014

IUBMB Life

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Human chorionic gonadotropin (hCG) is a glycoprotein produced by placental trophoblasts. Previous studies indicated that hCG could be responsible for the pregnancy-induced protection against breast cancer in women. It is reported that hCG decreases proliferation and invasion of breast cancer MCF-7 cells. Our research also demonstrates that hCG can reduce the proliferation of MCF-7 cells by downregulating the expression of proliferation markers, proliferating cell nuclear antigen (PCNA), and proliferation-related Ki-67 antigen (Ki-67). Interestingly, we find here that hCG elevates the state of cellular differentiation, as characterized by the upregulation of differentiation markers, b-casein, cytokeratin-18 (CK-18), and Ecadherin. Inhibition of hCG secretion or luteinizing hormone/ hCG receptors (LH/hCGRs) synthesis can weaken the effect of hCG on the induction of cell differentiation. Furthermore, hCG can suppress the expression of estrogen receptor alpha. hCG activated receptor-mediated cyclic adenosine monophosphate/ protein kinase A signaling pathway. These findings indicated that a protective effect of hCG against breast cancer may be associated with its growth inhibitory and differentiation induction function in breast cancer cells. V C 2014 IUBMB Life, 66(5): [352][353][354][355][356][357][358][359][360] 2014

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“…detection of morphine in rat tissues [10], demonstration of tetrahydrocannabinol [11] and phenobarbital [12] in mice tissues. However, the principle of antigen-antibody recognition can also be applied in histological specimens (immunohistochemistry), allowing detection of drugs in tissue sections.…”

Section: Introductionmentioning

confidence: 99%

Immunohistochemical demonstration of the amphetamine derivatives 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) in human post-mortem brain tissues and the pituitary gland

et al. 2003

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Abuse of amphetamine derivatives such as 3,4methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) is an important issue in current forensic practice and fatalities are not infrequent. Therefore, we investigated an immunohistochemical method to detect the amphetamine analogues MDMA and MDA in human tissues. For the staining procedure, the Catalysed Signal Amplification (CSA) method using peroxidase (HRP) provided by Dako ® and specific monoclonal antibodies were used. Appropriate controls for validation of the technique were included. The distribution of these designer drugs was studied in various brain regions including the four lobes, the basal ganglia, hypothalamus, hippocampus, corpus callosum, medulla oblongata, pons, cerebellar vermis and, additionally, in the pituitary gland. A distinct positive reaction was observed in all cortical brain regions and the neurons of the basal ganglia, the hypothalamus, the hippocampus and the cerebellar vermis but in the brainstem, relatively weak staining of neurons was seen. The reaction presented as a mainly diffuse cytoplasmic staining of the perikaryon of the neurons, and often axons and dendrites were also visualised. In addition, the immunoreactivity was present in the white matter. In the pituitary gland, however, distinct immunopositive cells were observed, with a prominent heterogeneity. The immunohistochemical findings were supported by the toxi-cological data. This immunostaining technique can be used as evidence of intake or even poisoning with MDMA and/or MDA and can be an interesting tool in forensic practice when the usual samples for toxicological analysis are not available. Furthermore, this method can be used to investigate the distribution of these substances in the human body.

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An Assessment of the Immunofluorescence Technique as a Method for Demonstrating the Histological Localization of Tetrahydrocannabinol in Mammalian Tissues

Cited by 5 publications

References 10 publications

Estrogen receptor α mediates proliferation of breast cancer MCF–7 cells via a p21/PCNA/E2F1‐dependent pathway

Estrogen receptor α mediates proliferation of breast cancer MCF–7 cells via a p21/PCNA/E2F1‐dependent pathway

Human chorionic gonadotropin decreases human breast cancer cell proliferation and promotes differentiation

Immunohistochemical demonstration of the amphetamine derivatives 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) in human post-mortem brain tissues and the pituitary gland

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