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A framework for the in vitro evaluation of cancer-relevant molecular characteristics and mitogenic potency of insulin analogues.

Baricevic I, Jones DR, Roberts DL, Lutzen A, Lundby A, Worm J, Hansen BF, Renehan AG - Carcinogenesis (2015)

Bottom Line: Accordingly, there is a regulatory mandate for cancer-related pre-clinical safety evaluation during insulin analogue development, but currently, there is no standardized framework for such in vitro evaluation.The meta-analysis, adding data from five additional studies, supported the hypothesis that ligand mitogenic potency, relative to human insulin, increases with increasing cell-specific IGF-IR/IR ratio.This study established a framework for the in vitro evaluation of cancer-relevant bioassays for comparisons of insulin analogues, and specifically consolidated earlier studies that determination of the cell-specific IGF-IR/IR ratio is crucial for the interpretation of ranking relative biological activities.

View Article: PubMed Central - PubMed

Affiliation: Faculty Institute of Cancer Sciences, Manchester Academic Health Science Centre, The Christie NHS Foundation Trust, University of Manchester, Wilmslow Road, Manchester M20 4BX, UK, Inositide Laboratory, Cancer Research UK Manchester Institute, Paterson Building, Manchester M20 4BX, UK and Diabetes Research Unit, Novo Nordisk A/S, Måløv, Denmark.

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Dose–response curves for increasing doses of the three ligands—insulin, X10 and IGF-I (a). For these experiments, we included zero ligand concentrations (data not shown)—there were no differences with effects for 1 pmol for any ligand, indicating no ‘background’ effect from the FCS. Each point in the figure represents the mean ± SD of a triplicate determination. The experiment was performed three times. From these curves, EC50 values were derived using GraphPad Prism and mitogenic potencies calculated relative to insulin as 100% (b). Regression line fitted to demonstrate the relationship between increasing IGF-IR/IR ratio and relative mitogenic potency. The ‘non-responder’ data for HT-29 are not included.
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Figure 5: Dose–response curves for increasing doses of the three ligands—insulin, X10 and IGF-I (a). For these experiments, we included zero ligand concentrations (data not shown)—there were no differences with effects for 1 pmol for any ligand, indicating no ‘background’ effect from the FCS. Each point in the figure represents the mean ± SD of a triplicate determination. The experiment was performed three times. From these curves, EC50 values were derived using GraphPad Prism and mitogenic potencies calculated relative to insulin as 100% (b). Regression line fitted to demonstrate the relationship between increasing IGF-IR/IR ratio and relative mitogenic potency. The ‘non-responder’ data for HT-29 are not included.

Mentions: Western blots (a) and densitometry (b) for expression of IR and IGF-IR in four cell lines: HCT 116, HT-29, COLO 205 and MCF7. IR and IGF-IR expression determined using the QIFIKIT method (c). The values for IGF-IR/IR ratio in (c) were used subsequently in Figures 5 and 6. SABC, specific antigen binding capacity.


A framework for the in vitro evaluation of cancer-relevant molecular characteristics and mitogenic potency of insulin analogues.

Baricevic I, Jones DR, Roberts DL, Lutzen A, Lundby A, Worm J, Hansen BF, Renehan AG - Carcinogenesis (2015)

Dose–response curves for increasing doses of the three ligands—insulin, X10 and IGF-I (a). For these experiments, we included zero ligand concentrations (data not shown)—there were no differences with effects for 1 pmol for any ligand, indicating no ‘background’ effect from the FCS. Each point in the figure represents the mean ± SD of a triplicate determination. The experiment was performed three times. From these curves, EC50 values were derived using GraphPad Prism and mitogenic potencies calculated relative to insulin as 100% (b). Regression line fitted to demonstrate the relationship between increasing IGF-IR/IR ratio and relative mitogenic potency. The ‘non-responder’ data for HT-29 are not included.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4552242&req=5

Figure 5: Dose–response curves for increasing doses of the three ligands—insulin, X10 and IGF-I (a). For these experiments, we included zero ligand concentrations (data not shown)—there were no differences with effects for 1 pmol for any ligand, indicating no ‘background’ effect from the FCS. Each point in the figure represents the mean ± SD of a triplicate determination. The experiment was performed three times. From these curves, EC50 values were derived using GraphPad Prism and mitogenic potencies calculated relative to insulin as 100% (b). Regression line fitted to demonstrate the relationship between increasing IGF-IR/IR ratio and relative mitogenic potency. The ‘non-responder’ data for HT-29 are not included.
Mentions: Western blots (a) and densitometry (b) for expression of IR and IGF-IR in four cell lines: HCT 116, HT-29, COLO 205 and MCF7. IR and IGF-IR expression determined using the QIFIKIT method (c). The values for IGF-IR/IR ratio in (c) were used subsequently in Figures 5 and 6. SABC, specific antigen binding capacity.

Bottom Line: Accordingly, there is a regulatory mandate for cancer-related pre-clinical safety evaluation during insulin analogue development, but currently, there is no standardized framework for such in vitro evaluation.The meta-analysis, adding data from five additional studies, supported the hypothesis that ligand mitogenic potency, relative to human insulin, increases with increasing cell-specific IGF-IR/IR ratio.This study established a framework for the in vitro evaluation of cancer-relevant bioassays for comparisons of insulin analogues, and specifically consolidated earlier studies that determination of the cell-specific IGF-IR/IR ratio is crucial for the interpretation of ranking relative biological activities.

View Article: PubMed Central - PubMed

Affiliation: Faculty Institute of Cancer Sciences, Manchester Academic Health Science Centre, The Christie NHS Foundation Trust, University of Manchester, Wilmslow Road, Manchester M20 4BX, UK, Inositide Laboratory, Cancer Research UK Manchester Institute, Paterson Building, Manchester M20 4BX, UK and Diabetes Research Unit, Novo Nordisk A/S, Måløv, Denmark.

Show MeSH
Related in: MedlinePlus