| RRC ID |
35777
|
| 著者 |
Inagawa Y, Yamada K, Yugawa T, Ohno S, Hiraoka N, Esaki M, Shibata T, Aoki K, Saya H, Kiyono T.
|
| タイトル |
A human cancer xenograft model utilizing normal pancreatic duct epithelial cells conditionally transformed with defined oncogenes.
|
| ジャーナル |
Carcinogenesis
|
| Abstract |
Pancreatic ductal adenocarcinomas (PDACs) are considered to arise through neoplastic transformation of human pancreatic duct epithelial cells (HPDECs). In order to evaluate the biological significance of genetic and epigenetic alterations in PDACs, we isolated primary HPDECs and established an in vitro carcinogenesis model. Firstly, lentivirus-mediated transduction of KRAS(G12V), MYC and human papillomavirus 16 (HPV16) E6/E7 under the control of a tetracyclin-inducible promoter efficiently immortalized and transformed primary HPDECs, which gave rise to adenocarcinomas subcutaneously in an immune-deficient mouse xenograft model, depending on expression of the four genes. The tumors regressed promptly upon shutting-off the oncogenes, and the remaining tissues showed histological features corresponding to normal ductal structures with simple columnar epithelium. Reexpression of the oncogenes resulted in development of multiple PDACs through pancreatic intraepithelial neoplasia-like structures. We also succeeded in efficient immortalization of primary HPDECs with transduction of mutant CDK4, cyclin D1 and TERT. The cells maintained a normal diploid status and formed duct-like structures in a three-dimensional culture. In combination with p53 silencing, KRAS(G12V) alone was sufficient to fully transform the immortalized HPDECs, and MYC markedly accelerated the development of tumors. Our PDAC model supports critical roles of KRAS mutations, inactivation of the p53 and p16-pRB pathways, active telomerase and MYC expression in pancreatic carcinogenesis and thus recapitulates many features of human PDAC development. The present system with reversible control of oncogene expression enabled de novo development of PDAC from quasinormal human tissues preformed subcutaneously in mice and might be applicable to carcinogenesis models in many organ sites.
|
| 巻・号 |
35(8)
|
| ページ |
1840-6
|
| 公開日 |
2014-8-1
|
| DOI |
10.1093/carcin/bgu112
|
| PII |
bgu112
|
| PMID |
24858378
|
| MeSH |
Animals
Blotting, Western
Carcinoma, Pancreatic Ductal / genetics
Carcinoma, Pancreatic Ductal / metabolism
Carcinoma, Pancreatic Ductal / pathology*
Cell Culture Techniques
Cell Transformation, Neoplastic / genetics
Cell Transformation, Neoplastic / metabolism
Cell Transformation, Neoplastic / pathology*
Cells, Cultured
Cyclin D1 / genetics
Cyclin-Dependent Kinase 4 / genetics
Epithelial Cells / metabolism
Epithelial Cells / pathology*
Female
Gene Expression Regulation, Neoplastic*
Humans
Immunoenzyme Techniques
Mice
Mice, Inbred BALB C
Mice, Nude
Mutation / genetics
Oncogenes / physiology*
Pancreatic Ducts / metabolism
Pancreatic Ducts / pathology*
Pancreatic Neoplasms / genetics
Pancreatic Neoplasms / metabolism
Pancreatic Neoplasms / pathology*
Proto-Oncogene Proteins / genetics
Proto-Oncogene Proteins p21(ras)
RNA, Messenger / genetics
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Telomerase / genetics
Tumor Suppressor Protein p53 / genetics
Xenograft Model Antitumor Assays
ras Proteins / genetics
|
| IF |
4.603
|
| 引用数 |
9
|
|
WOS 分野
|
ONCOLOGY
|
| リソース情報 |
| 遺伝子材料 |
CSII-CMV-RfA (RDB04386)
CSII-EF-RfA (RDB04387). |
