Characterization of muscarinic receptors in the human melanoma cell line SK-Mel-28 via calcium mobilization

Noda, S.; Lammerding–Köppel, Maria; Oettling, G.; Drews, Ulrich

doi:10.1016/s0304-3835(98)00215-8

Cited by 30 publications

(27 citation statements)

References 15 publications

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“…The intracellular calcium mobilization after muscarinic stimulation was similar to the reaction curves in embryonic cells [Oettling et al, 1992] and in the human melanoma cell line Sk-mel 28 [Noda et al, 1998]. The effective doses were also in the same range.…”

Section: Discussionsupporting

confidence: 72%

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Identification and Functional Characterization of the Muscarinic Receptor M3 in the Human Keratinocyte Cell Line HaCaT

Metzger

Just²,

Boss³

et al. 2005

Cells Tissues Organs

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In the epidermis muscarinic cholinergic receptors are expressed in keratinocytes during normal differentiation and in regeneration of the skin. The muscarinic receptors were implicated to regulate proliferation, migration and differentiation of keratinocytes. Therefore, we characterized the muscarinic receptor in the spontaneously immortalized human keratinocyte cell line HaCaT and studied its role in cell migration. By RT-PCR, Western blot and immunocytochemistry we show that HaCaT cells express the M3 receptor subtype. This receptor was further characterized by fluorometric measurement of calcium mobilization in cell suspensions. After muscarinic stimulation a peak reaction of intracellular calcium mobilization occurred followed by a plateau phase of extracellular influx which was reversible by atropine. In dose-response curves the effective concentrations of carbachol and acetylcholine were determined as 40 and 1 µM, respectively. The migratory activity of HaCaT cells was studied with the phagokinetic track assay. Treatment with the muscarinic agonist carbachol had no effect on cell migration. However, carbachol diminished the EGF-induced increase in migratory activity by about 26%. Interestingly, EGF and carbachol had additive effects on calcium mobilization.

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Section: Discussionsupporting

confidence: 72%

“…After equilibration acetylcholine or carbachol was added to stimulate the mAChRs. An increase in cytoplasmic Ca 2+ causes a decrease of fl uorescence intensity [Noda et al, 1998]. Effective dose (ED 50 ) is the agonist concentration yielding 50% of the maximal effect.…”

Section: Fluorometric Calcium Measurementsmentioning

confidence: 99%

Identification and Functional Characterization of the Muscarinic Receptor M3 in the Human Keratinocyte Cell Line HaCaT

Metzger

Just²,

Boss³

et al. 2005

Cells Tissues Organs

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“…These receptors have all been cloned and well characterized (152,285,357). Muscarinergic receptors have been detected in melanoma cell lines in vitro (446,588), keratinocytes (285, 287, 288, 290), fibroblasts (123), and endothelial cells (308). In rat skin, the m 2 receptor was detected in nerve fibers (308) where it may be involved in pain and nociception (798).…”

Section: Ach Receptorsmentioning

confidence: 99%

Neuronal Control of Skin Function: The Skin as a Neuroimmunoendocrine Organ

Roosterman¹,

Goerge

Schneider

et al. 2006

Physiological Reviews

548

521

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This review focuses on the role of the peripheral nervous system in cutaneous biology and disease. During the last few years, a modern concept of an interactive network between cutaneous nerves, the neuroendocrine axis, and the immune system has been established. We learned that neurocutaneous interactions influence a variety of physiological and pathophysiological functions, including cell growth, immunity, inflammation, pruritus, and wound healing. This interaction is mediated by primary afferent as well as autonomic nerves, which release neuromediators and activate specific receptors on many target cells in the skin. A dense network of sensory nerves releases neuropeptides, thereby modulating inflammation, cell growth, and the immune responses in the skin. Neurotrophic factors, in addition to regulating nerve growth, participate in many properties of skin function. The skin expresses a variety of neurohormone receptors coupled to heterotrimeric G proteins that are tightly involved in skin homeostasis and inflammation. This neurohormone-receptor interaction is modulated by endopeptidases, which are able to terminate neuropeptide-induced inflammatory or immune responses. Neuronal proteinase-activated receptors or transient receptor potential ion channels are recently described receptors that may have been important in regulating neurogenic inflammation, pain, and pruritus. Together, a close multidirectional interaction between neuromediators, high-affinity receptors, and regulatory proteases is critically involved to maintain tissue integrity and regulate inflammatory responses in the skin. A deeper understanding of cutaneous neuroimmunoendocrinology may help to develop new strategies for the treatment of several skin diseases.

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“…Amplification of cholinesterase genes occurs in ovarian tumors and leukemias, as well as in pre-malignant conditions, such as polycythemia vera, where a 15-fold increase in AChE and butyrylcholinesterase genes was observed [2]. The abundant presence of cholinesterase activity, cholinesterase proteins and RNA messages, and acetylcholine receptors in many tumor cells [3][4][5][6] suggests that cholinergic signaling may be functionally important.…”

Section: Introductionmentioning

confidence: 99%

Acetylcholinesterase supports anchorage independence in colon cancer

Syed

Fenoglio‐Preiser

Skau

et al. 2008

Clin Exp Metastasis

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Various roles have been attributed to Acetylcholinesterase (AChE) in cancer. Evidence exists for a pro-apoptotic function, consistent with a protective role of AChE. Because other reports suggested that upregulated AChE in some tumors may control cell adhesion, we tested the effects of AChE on anchorage independence (an essential component of metastasis) of colon tumor cells. Several AChE inhibitors dose-dependently suppressed colony formation of HTB-38 cells in soft agar. This effect of AChE was confirmed with HTB-38 cells stably overexpressing AChE. In contrast, cell proliferation was not altered by the effective doses of these chemical inhibitors or by transfected AChE. Protection from cell cycle arrest consecutive to cancer cell detachment may be conveyed by changes in cell-matrix interactions. Reflective of such changes, the AChE overexpressing cells adhered more strongly to Fibronectin than did the vector controls. The AChE-dependent adhesion was RGD-dependent and accompanied by increased c-Myb DNA-binding, suggesting that AChE upregulates an Integrin receptor via c-Myb. In support of these observations, we find AChE message and protein to be expressed in a large fraction of colon cancers and in all colon tumor cell lines analyzed, but only rarely in normal colon specimens. Our results imply a dual role for AChE in colon cancer. While the anti-apoptotic effects of AChE may be protective against early stages of tumorigenesis, this gene product may support the later stages of transformation by enhancing anchorage independent growth. The induction of Integrins could render the cells independent of microenvironmental cues and override cell cycle arrest after deadhesion.

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Characterization of muscarinic receptors in the human melanoma cell line SK-Mel-28 via calcium mobilization

Cited by 30 publications

References 15 publications

Identification and Functional Characterization of the Muscarinic Receptor M3 in the Human Keratinocyte Cell Line HaCaT

Identification and Functional Characterization of the Muscarinic Receptor M3 in the Human Keratinocyte Cell Line HaCaT

Neuronal Control of Skin Function: The Skin as a Neuroimmunoendocrine Organ

Acetylcholinesterase supports anchorage independence in colon cancer

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