Hallo,
ich bin auf der Suche nach zusätzlichen Optionen über Methadon gestolpert. Kürzlich gab es dazu die folgende Veröffentlichung. Wie ist diese einzuordnen? Danke.
Der Tunel Assay bestätigt den durch Methadon induzierten Zelltod in Glioblastomzelllinien und normalen Astrozyten
Deutsche Gesellschaft für Neurochirurgie. 72. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), Joint Meeting mit der Polnischen Gesellschaft für Neurochirurgie.
Veröffentlicht: 4. Juni 2021
Objective:
Glioblastoma (GBM) is the most frequent primary brain tumor in the adulthood and carries a dismal prognosis. Recently, it has been shown, that D,L-methadone induced apoptosis in GBM cell lines detected by annexin V FACS analysis. To confirm and to assess the executive state of apoptotic cell death with another method, we applied TUEL labelling assay.
Methods:
We utilized three different established rat GBM cell lines (9L, F98, S635) and a normal rat astrocytes as a primary culture from Fisher 344 rats. All cell lines were exposed to 5 concentrations of D,L-methadone (0.3, 1.0, 15, 30, 45 µg/ml) alone or in addition to TMZ (100 µM). After 6 days cells were harvested and apoptosis was detected by annexin V FACS analysis and fragmentated DNA was detected by TUNEL labelling assay to confirm apoptosis.
Results:
All tested cell lines express of the μ-opioid receptor was similarly; in particular, there was no difference between the GBM cells compared to normal astrocytes. Apoptosis, detected by annexin V FACS, is induced in all cell lines by D,L-methadone alone beginning at 15µg/ml and in addition to TMZ with 0.3µg/ml (9L), respectively with 15µg/ml D,L-methadone (F98, S635, 9L). The tunnel assay showed cell death with defragmented DNA in all cell lines, conforming apoptosis detected by annexin V FACS analysis. The TUNEL labelling experiments confirmed the data of apoptosis. In comparison to GBM cell lines, the astrocytes react refractory against D,L-methadone alone, confirmed in apoptosis and TUNEL labelling assay.
Conclusion:
TUNEL labelling experiments confirmed the findings of annexiv V FACS, indicating the induction of apoptotic cell death in higher concentrations of methadone (> 15 mg/ml), with the astrocytes showing the lowest susceptibility.