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Volume 40 Issue 6

Jun. 2024

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Article Contents

Article Navigation > Journal of Clinical Hepatology > 2024 > 40(6): 1231-1239

Establishment and in vivo imaging observation of a nude mouse model of type 2 diabetes mellitus and pancreatic cancer

DOI: 10.12449/JCH240625

1.
Department of Pathology，Wuming Hospital Affiliated to Guangxi Medical University，Nanning 530199，China
2.
Department of Pathology，The Second Affiliated Hospital of Guangxi Medical University，Nanning 530005，China
3.
Department of Laboratory Medicine，The First Affiliated Hospital of Guangxi Medical University，Nanning 530021，China

Research funding:

National Natural Science Foundation of China (81860508);

Guangxi Medical and Health Appropriate Technology Development and Promotion Project (S2018040);

Guangxi Health and Family Planning Commission Self-raised Scientific Research Projects (Z20180950)

More Information

Corresponding author: QIN Wen， fenglingcao1980@163.com （ORCID： 0009-0002-8282-7388）
Received Date: 2023-10-17
Accepted Date: 2023-12-26
Published Date: 2024-06-25

Abstract

Objective To establish a nude mouse model of type 2 diabetes mellitus （T2DM） and pancreatic cancer that allows dynamic observation of tumor formation process and facilitates invivo research. Methods At first， human pancreatic cancer PANC-1 cells were transfected with lentiviral vector GV260 to construct the pancreatic cancer cell line PANC-1-Luc with stable expression of firefly luciferase. Then， 36 specific pathogen-free nude mice were randomly divided into control group with 12 mice and model group with 24 mice （nude mice with T2DM and pancreatic cancer）. The mice in the control group were fed with breeding diet and were then given ectopic subcutaneous implantation of PANC-1-Luc cells， and those in the model group were first given high-fat diet and intraperitoneal injection of 1% STZ， followed by ectopic subcutaneous implantation of PANC-1-Luc cells. The fluorescence invivo imaging system and the manual measurement method were used for simultaneous and dynamic monitoring of the growth of pancreatic cancer in nude mice in the two groups， and the tumor growth curve was plotted to investigate the correlation between fluorescence value and tumor volume. Subcutaneous tumors and pancreatic islets were observed under a microscope to verify whether the model was successfully established， and immunohistochemistry was used to measure the expression of Ki-67 in tumor tissue to investigate the influence of hyperglycemia on the growth of pancreatic cancer in nude mice. The independent-samples t test was used for comparison of normally distributed continuous data between groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups. Results The optimal virus titer was determined as 5×10⁷ TU/mL for the stable transfection of lentiviral vector in PANC-1 cells， and the optimal concentration selected with puromycin was 20 μg/mL， with an optimal selection time of 9 days. The fluorescence value of PANC-1-Luc cells was linearly and positively correlated with the number of cells， with the linear equation of y=42.56x－42 504 （r=0.977， P=0.004）. The blood glucose value of T2DM nude mice was 23.05 （19.25‍ — ‍26.40） mmol/L， with a blood glucose level of >11.1 mmol/L in each nude mouse， and there was a significant difference in blood glucose value between the T2DM nude mice and the control nude ［6.15 （5.20‍ — ‍7.30） mmol/L］（Z=-8.45， P<0.001）. Compared with the control group， the model group had reductions in the number and volume of pancreatic islets， with irregular shapes and unclear boundaries， and pathological examination confirmed that the xenograft tumor was pancreatic cancer tissue， which showed that the model was established successfully. In the model group， there was a linear positive correlation between subcutaneous tumor size and fluorescence values， with the linear equation of y=232 348 691x－8 258 608 （r=0.911， P=0.031）. The model group had a significantly higher positive rate of Ki-67 than the control group （50.333%± 7.808% vs 15.917%±4.055%， t=13.55， P<0.001）， suggesting rapid tumor proliferation in the model group. Conclusion The T2DM nude mouse model of pancreatic cancer established in this study can simulate the pathological process of the development and progression of pancreatic cancer in the context of T2DM and dynamically observe the influence of hyperglycemia on the growth of pancreatic cancer cells invivo， thereby providing a new experimental vector for the invivo study of the development and progression of pancreatic cancer in the context of T2DM.
- Diabetes Mellitus， Type 2,
- Pancreatic Neoplasms,
- Disease Models， Animal

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