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Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells.

Kerros C, Cavey T, Sola B, Jauzac P, Allouche S - J. Exp. Clin. Cancer Res. (2009)

Bottom Line: opioid and somatostatin receptors (SSTRs) that can assemble as heterodimer were individually reported to modulate malignant cell proliferation and to favour apoptosis.XTT assays and cell cycle studies provide no evidence for a significant effect upon opioid or somatostatin receptors stimulation.Furthermore, neither direct effect nor potentiation of the Fas-receptor pathway was detected on apoptosis after these treatments. these data suggest that SSTRs or opioid receptors expression is not a guaranty for an anti-tumoral action in U266 cell line.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de biologie moléculaire et cellulaire de la signalisation, UPRES-EA 3919, IFR 146 ICORE, Université de Caen, Caen, France. ckerros@yahoo.fr

ABSTRACT

Background: opioid and somatostatin receptors (SSTRs) that can assemble as heterodimer were individually reported to modulate malignant cell proliferation and to favour apoptosis.

Materials and methods: SSTRs and opioid receptors expression were examined by RT-PCR, western-blot and binding assays, cell proliferation was studied by XTT assay and propidium iodide (PI) staining and apoptosis by annexin V-PI labelling.

Results: almost all human malignant haematological cell lines studied here expressed the five SSTRs. Further experiments were conducted on the human U266 multiple myeloma cells, which express also micro-opioid receptors (MOP-R). XTT assays and cell cycle studies provide no evidence for a significant effect upon opioid or somatostatin receptors stimulation. Furthermore, neither direct effect nor potentiation of the Fas-receptor pathway was detected on apoptosis after these treatments.

Conclusion: these data suggest that SSTRs or opioid receptors expression is not a guaranty for an anti-tumoral action in U266 cell line.

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Cell cycle distribution of U266 cells after SSTR stimulation. Exponentially growing cells were incubated with 10 μM Sst or Oct, or with 0.1 mg/mL 7C11 (agonistic Fas antibody) for 72 h. DNA content analysis was done after PI staining of ethanol-permeabilized cells. % of each cell cycle phase are summarized in the Table 2. Data shown are representative of 3 independent experiments.
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Figure 3: Cell cycle distribution of U266 cells after SSTR stimulation. Exponentially growing cells were incubated with 10 μM Sst or Oct, or with 0.1 mg/mL 7C11 (agonistic Fas antibody) for 72 h. DNA content analysis was done after PI staining of ethanol-permeabilized cells. % of each cell cycle phase are summarized in the Table 2. Data shown are representative of 3 independent experiments.

Mentions: We confirmed by using an alternative method, that SSTR agonists were ineffective to regulate U266 cell proliferation. Distribution in the cell cycle of control or agonist-pretreated U266 cells was determined after PI staining by flow cytometry. A low (10 nM) or a high concentration (10 μM) of Sst or Oct alone, or in combination with Css were selected and cells were exposed during 24, 48 or 72 h. A representative experiment is depicted in the Figure 3 showing that neither Sst (10 μM) nor Oct (10 μM) were able to promote changes in cell cycle distribution compared to control cells after 72 h. Similar data were obtained for 24 and 48 h pretreatment (data not shown). The percentage of each phase was determined for control or agonist-pretreated cells and these data are summarised in the Table 3.


Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells.

Kerros C, Cavey T, Sola B, Jauzac P, Allouche S - J. Exp. Clin. Cancer Res. (2009)

Cell cycle distribution of U266 cells after SSTR stimulation. Exponentially growing cells were incubated with 10 μM Sst or Oct, or with 0.1 mg/mL 7C11 (agonistic Fas antibody) for 72 h. DNA content analysis was done after PI staining of ethanol-permeabilized cells. % of each cell cycle phase are summarized in the Table 2. Data shown are representative of 3 independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2698864&req=5

Figure 3: Cell cycle distribution of U266 cells after SSTR stimulation. Exponentially growing cells were incubated with 10 μM Sst or Oct, or with 0.1 mg/mL 7C11 (agonistic Fas antibody) for 72 h. DNA content analysis was done after PI staining of ethanol-permeabilized cells. % of each cell cycle phase are summarized in the Table 2. Data shown are representative of 3 independent experiments.
Mentions: We confirmed by using an alternative method, that SSTR agonists were ineffective to regulate U266 cell proliferation. Distribution in the cell cycle of control or agonist-pretreated U266 cells was determined after PI staining by flow cytometry. A low (10 nM) or a high concentration (10 μM) of Sst or Oct alone, or in combination with Css were selected and cells were exposed during 24, 48 or 72 h. A representative experiment is depicted in the Figure 3 showing that neither Sst (10 μM) nor Oct (10 μM) were able to promote changes in cell cycle distribution compared to control cells after 72 h. Similar data were obtained for 24 and 48 h pretreatment (data not shown). The percentage of each phase was determined for control or agonist-pretreated cells and these data are summarised in the Table 3.

Bottom Line: opioid and somatostatin receptors (SSTRs) that can assemble as heterodimer were individually reported to modulate malignant cell proliferation and to favour apoptosis.XTT assays and cell cycle studies provide no evidence for a significant effect upon opioid or somatostatin receptors stimulation.Furthermore, neither direct effect nor potentiation of the Fas-receptor pathway was detected on apoptosis after these treatments. these data suggest that SSTRs or opioid receptors expression is not a guaranty for an anti-tumoral action in U266 cell line.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de biologie moléculaire et cellulaire de la signalisation, UPRES-EA 3919, IFR 146 ICORE, Université de Caen, Caen, France. ckerros@yahoo.fr

ABSTRACT

Background: opioid and somatostatin receptors (SSTRs) that can assemble as heterodimer were individually reported to modulate malignant cell proliferation and to favour apoptosis.

Materials and methods: SSTRs and opioid receptors expression were examined by RT-PCR, western-blot and binding assays, cell proliferation was studied by XTT assay and propidium iodide (PI) staining and apoptosis by annexin V-PI labelling.

Results: almost all human malignant haematological cell lines studied here expressed the five SSTRs. Further experiments were conducted on the human U266 multiple myeloma cells, which express also micro-opioid receptors (MOP-R). XTT assays and cell cycle studies provide no evidence for a significant effect upon opioid or somatostatin receptors stimulation. Furthermore, neither direct effect nor potentiation of the Fas-receptor pathway was detected on apoptosis after these treatments.

Conclusion: these data suggest that SSTRs or opioid receptors expression is not a guaranty for an anti-tumoral action in U266 cell line.

Show MeSH
Related in: MedlinePlus