The Heterogeneity of Response of PC12 Cells from Different Laboratories to Nerve Growth Factor and Pituitary Adenylate Cyclase-Activating Polypeptide Questions the Reproducibility of Studies Carried Out with Tumor Cell Lines
Abstract:<b><i>Background:</i></b> PC12 pheochromocytoma tumor cell lines are widely used to decipher the intracellular signaling mechanisms mediating the effects of some growth factors. Nevertheless, the disparity in appearance of some PC12 cell lines used in the different publications questions our ability to compare the results obtained by the numerous laboratories which use them. This led us to analyze the phenotypic aspect and transcriptomic expression of 5 PC12 cell lines from different or… Show more
“…To be complete, one should mention other non-cancer cells used in cell biology studies, particularly CHO cells, an aneuploid hamster-derived epithelial cell line. Even though this cell line has been very carefully analyzed [40], it remains that the multitude of clones derived are slightly different from each other [41,42], leading to a similar view to PC12 cancer cells, for which clones have widely different expression profiles, leading to completely different conclusions in GPCR pathways [43]. Therefore, the suitability of these host cell lines for the study of "physiological-like" conditions should be questioned.…”
That signaling bias is a nth level of complexity in the understanding of G protein-coupled receptor (GPCR) activation is a first fact. That its exhaustive description, including the mode d’emploi of its quantitative measurement, remains a challenge is a second fact. That the use of this concept is promising for the design of drug candidates is a third fact. That the translation of signaling biases observed into in vivo specific effects is well documented is a fourth fact. However, the road to apply those aspects of receptology to a systematic description of a ligand and, a fortiori, of a drug candidate, still necessitates a huge body of studies. In the present commentary, the merits of the molecular description of receptor bias signaling are highlighted and the ligand induced-fit impact on GPCR structure, as well as on the functional repertoire of GPCRs, is discussed. An emphasis is given to the practical aspects during drug design, and, thus, the practical limitations of the current approaches, particularly in the context of as soon as the data are transferred to more integrated/living systems, might be a major limitation.
“…To be complete, one should mention other non-cancer cells used in cell biology studies, particularly CHO cells, an aneuploid hamster-derived epithelial cell line. Even though this cell line has been very carefully analyzed [40], it remains that the multitude of clones derived are slightly different from each other [41,42], leading to a similar view to PC12 cancer cells, for which clones have widely different expression profiles, leading to completely different conclusions in GPCR pathways [43]. Therefore, the suitability of these host cell lines for the study of "physiological-like" conditions should be questioned.…”
That signaling bias is a nth level of complexity in the understanding of G protein-coupled receptor (GPCR) activation is a first fact. That its exhaustive description, including the mode d’emploi of its quantitative measurement, remains a challenge is a second fact. That the use of this concept is promising for the design of drug candidates is a third fact. That the translation of signaling biases observed into in vivo specific effects is well documented is a fourth fact. However, the road to apply those aspects of receptology to a systematic description of a ligand and, a fortiori, of a drug candidate, still necessitates a huge body of studies. In the present commentary, the merits of the molecular description of receptor bias signaling are highlighted and the ligand induced-fit impact on GPCR structure, as well as on the functional repertoire of GPCRs, is discussed. An emphasis is given to the practical aspects during drug design, and, thus, the practical limitations of the current approaches, particularly in the context of as soon as the data are transferred to more integrated/living systems, might be a major limitation.
Background:
Nepeta binaludensis Jamzad is a perennial medicinal plant that exhibits various pharmacological effects. However, the neuroprotective effects of Nepeta binaludensis extracts have not yet been investigated. This study aimed to examine the effects of N. binaludensis hydro-alcoholic extract (NBE) on oxidative stress markers and apoptosis-related proteins in PC12 cells exposed to oxygen-glucose deprivation/reperfusion (OGD/R).
Methods:
PC12 cells were pretreated with NBE (at concentration range of 10-200 μg/ml) before exposure to OGD condition for 6 h followed by a 24 h reoxygenation. Cell viability, the production of the reactive oxygen species (ROS), lipid peroxidation (LPO), and the levels of apoptosis-related proteins were evaluated using MTT, fluorimetry, and western blot analysis, respectively.
Results:
Survival of the cells preincubated for 6 h with NBE increased to 90.20 ± 15.62% compared with those subjected to OGD/R alone (51.26 ± 7.77 %, p < 0.001). ROS formation was also decreased following incubation with 200 µg/ml of NBE to 125.3 ± 18.38% compared to OGD/R group (356.9 ± 70.48%, p < 0.001). LPO was also suppressed after incubation with NBE to 155.5 ± 21.21% compared to the OGD/R group (260.5 ± 9.727%, p<0.001). NBE restored Bax/Bcl-2 ratio (1.3-fold of control), and cleaved caspase-3 (1.58-fold of control, p < 0.001).
Conclusion:
These results suggest that NBE may offer neuroprotection properties against OGD/R-induced toxicity through modulation of oxidative stress and apoptotic responses.
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