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Isolating and Analyzing Cells of the Pancreas Mesenchyme by Flow Cytometry
Published on: January 28, 2017
Gene Expression Analysis of Early Stage Changes in Pancreatic Cancer by KrasG12D Transfer in Pancreatic
1Department of Nutrition and Food Sciences, Mukogawa Women's University, Nishinomiya, Hyogo, Japan.
Objective:
The aim of this study is to elucidate the function of Kras, which is involved in the development and progression of pancreatic cancer.
Methods:
We have previously isolated pancreas-derived cells from mice and induced genes expressed in pancreatic ducts and exocrine and endocrine cells by expressing transcription factor genes and a plasmid expressing polyoma ori to create cells with stable foreign gene expression. Comprehensive gene analysis was used to analyze the function of Kras by introducing wild-type Kras and mutant Kras into these cells.
Results:
No changes in cell morphology were observed with wild-type Kras expression, but expression of mutant Kras led to cystic changes. Mutant Kras gene transfer induced the expression of Mus5ac and Cck genes. Gene expression in these cells was detected by microarray analysis, and comparison of gene profiles showed that the genes promoted amoeboid cell assembly and immunosuppression.
Conclusion:
The use of these cells will facilitate screening of drugs that block Kras function and elucidate how Kras induces pancreatic cancer.
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