A reporter system evaluates tumorigenesis, metastasis, β-catenin/MMP regulation, and druggability
| Autor: | Ken ichi Kozaki, Kuniaki Okamoto, Kisho Ono, Kazumi Ohyama, Yusaku Hamada, Masaharu Takigawa, Motoharu Iizuka, Norio Sogawa, Yuka Okusha, Chiharu Sogawa, Ryu Kawasaki, Takanori Eguchi |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Carcinogenesis
0206 medical engineering Biomedical Engineering Druggability Artesunate Bioengineering 02 engineering and technology MMP9 medicine.disease_cause Biochemistry Dexamethasone Fluorescence Metastasis Biomaterials 03 medical and health sciences syngeneic transplantation Genes Reporter Cell Line Tumor cancer metastasis medicine Animals Humans 3D tumoroid reporter assay RNA Messenger Neoplasm Metastasis Promoter Regions Genetic Transcription factor beta Catenin 030304 developmental biology 0303 health sciences Mice Inbred BALB C Chemistry Protein Stability Cancer medicine.disease tumoroid (tumor organoid) 020601 biomedical engineering Transplantation body regions Matrix Metalloproteinase 9 Gelatinases Catenin Colonic Neoplasms Cancer research Tetradecanoylphorbol Acetate Female metalloproteinase Wnt/β-catenin signaling |
| Zdroj: | Tissue Engineering Part A.. 25(19-20):1413-1425 |
| ISSN: | 1937-3341 |
| Popis: | Cancer invasion, metastasis, and therapy resistance are the crucial phenomena in cancer malignancy. The high-expression of matrix metalloproteinase 9 (MMP9) is a biomarker as well as a causal factor of cancer invasiveness and metastatic activity. However, a regulatory mechanism underlying MMP9 expression in cancer is not clarified yet. Additionally, a new strategy for anti-cancer drug discovery is becoming an important clue. In the present study, we aimed (i) to develop a novel reporter system evaluating tumorigenesis, invasiveness, metastasis, and druggability with a combination of three-dimensional (3D) tumoroid model and Mmp9 promoter and (ii) to examine pharmacological actions of anti-cancer medications using this reporter system. High expression and genetic amplification of MMP9 were found in colon cancer cases. We found that proximal promoter sequences of MMP9 in murine and human contained conserved binding sites for transcription factors β-catenin/TCF/LEF, glucocorticoid receptor (GR), and NF-κB. The murine Mmp9 promoter (-569 to +19) was markedly activated in metastatic colon cancer cells and additionally activated by tumoroid formation and by β-catenin signaling stimulator lithium chloride (LiCl). The Mmp9 promoter-driven fluorescent reporter cells enabled the monitoring of activities of MMP9/gelatinase, tumorigenesis, invasion, and metastasis in allogeneic/syngenic transplantation experiments. We also demonstrated pharmacological actions as follows. ids Dexamethasone and hydrocortisone, steroidal medications binding to GR, inhibited the Mmp9 promoter but did not inhibit tumorigenesis. On the other hand, an antimetabolite 5-fluorouracil, a golden standard for colon cancer chemotherapy, inhibited tumoroid formation but did not inhibit Mmp9 promoter activity. Notably, anti-malaria medication artesunate inhibited both tumorigenesis and the Mmp9 promoter in vitro, potentially through inhibition of β-catenin/TCF/LEF signaling. Thus, this novel reporter system enabled monitoring tumorigenesis, invasiveness, metastasis, key regulatory signalings such as β-catenin/MMP9 axis, and druggability. |
| Databáze: | OpenAIRE |
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