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Characterization and comparison of insulinoma tumor model and pancreatic damage caused by the tumor, and identification of possible markers

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Abstract

Insulinoma is a neuroendocrine tumor. It arises from the uncontrolled proliferation of pancreatic β cells. In this study, we created an insulinoma tumor model in nude mice. INS-1 cells were injected in two different ways, subcutaneously (S.C.) or intraperitoneally (I.P.). Body weight, tumor weight, and size were measured. ELISA kits were used analyze to Glucose, insulin, and CA19-9 levels in serum, pancreas, and tumor tissues. KCNN4, KCNK1, GLUT2, IR, HSP70, HSF1, and HSP90 levels were analyzed by western blotting of membrane and/or cytosolic fractions of tumor and pancreas tissue. Tumor formation occurred in nude mice, but it did not occur in Wistar albino rats. The tumor has neuroendocrine cell morphology. Insulin and CA19-9 levels increased in pancreas tissue. In tumor tissue, KCNN4 levels were higher in both membrane and cytosolic fractions, while KCNK1 levels were lower in the membrane fraction of the S.C. group. HSP70 levels were also lower in the S.C. group. In pancreas tissue, KCNK1 levels were lower in the membrane fraction of the S.C. and I.P. groups. GLUT2 levels increased in both groups according to the control group, while IR levels decreased in the S.C. group compared to the control group. However, HSF1 levels increased in the I.P. group, while HSP90 decreased in the S.C. group in pancreatic tissues. The S.C. group is a more suitable insulinoma tumor model. KCNN4, KCNK1, and HSP70 proteins may be important biomarkers in the diagnosis and treatment of insulinoma.

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Data availability

The data generated during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors would like to thank Prof. Claes B. Wollheim (University Medical Center, Geneva) for providing the kind gift of insulinoma INS-1 cell lines. The authors would like to thank Prof. Dr. Erhan Aysan (Yeditepe University Hospital, Department of General Surgery, Endocrine Surgery Unit, Istanbul, Turkey) for his valuable information on endocrine tumor formation.

Funding

This work was supported by the Scientific Research Project Coordination Unit of Istanbul University. Project No: FBA-29893.

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Authors and Affiliations

  1. Faculty of Science, Department of Biology, Istanbul University, 34134- Vezneciler, Istanbul, Turkey

    Ayse Karatug Kacar & Sehnaz Bolkent

  2. School of Medicine, Department of Pathology, Koç University, Istanbul, Turkey

    Pinar Bulutay

  3. Center for Immunology and Inflammation, Feinstein Institutes for Medical Research, Manhasset, NY, USA

    Dilara Aylar

  4. Institute of Health Science, Department of Anatomy, Medipol University, Istanbul, Turkey

    Mert Celikten

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Contributions

A.K.K., created the main idea, experimental design, all cell culture experiments done, control of animals during experimentation, taking of tissue, and tumor samples, determination of insulin and CA-19-9 levels, preparation of tissue and tumor samples, determinate of proteins by western blotting, evaluation of all experimental results, done statistics, written manuscript. P.B., Pathological examination of tissue and tumor samples taken at the end of the experiment and immunohistochemically marking of the samples with insulin. D.A., cell culture experiments, control animals during experimentation and label proteins by western blotting. M. C., INS-1 cell injection to experimental animals (rats and nude mice), control of animals during the experiment and termination of the experiment, taken of blood, tissue, and tumor samples. S.B., contributed to providing the necessary facilities for conducting the experiments and edited the manuscript.

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Correspondence to Ayse Karatug Kacar.

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The authors declare no competing interests.

Ethical approval and consent to Participate

This article contains studies with animals performed. This experimental study was carried out at the Experimental Animals Laboratory of Bezmialem University, in 2019–2021. The project was approved by the local ethics committee for animal experiments at Bezmialem University (2017/243).

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Karatug Kacar, A., Bulutay, P., Aylar, D. et al. Characterization and comparison of insulinoma tumor model and pancreatic damage caused by the tumor, and identification of possible markers. Mol Biol Rep 51, 109 (2024). https://doi.org/10.1007/s11033-023-08942-z

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