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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Jul 31, 2018 |
| Title |
Identification of selective inhibitors for diffuse-type gastric cancer cells by screening of annotated compounds in E-cadherin/p53 double conditional knockout mice |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Diffuse-type gastric cancer (DGC) exhibits rapid disease progression and a poor patient prognosis. We have previously established an E-cadherin/p53 double conditional knockout (DCKO) mouse line as the first genetically engineered one, which morphologically and molecularly recapitulates human DGC. In this study, we explored low-molecular-weight drugs selectively eliminating mouse DGC cells, and then validated their inhibitory effects on human DGC. We first derived mouse gastric cancer (GC) cell lines from DGC of the DCKO mice, which notably demonstrated enhanced tumorigenic activity in immunodeficient mice and acquired tolerance to cytotoxic anti-cancer agents. We next performed a synthetic lethal screening of 1535 annotated chemical compounds by using them. Comparing cell viability of the E-cadherin/p53-deficient GC and p53-deficient gastric epithelial (GE) cells under treatment with the compound library, we identified 27 candidates with specific toxicity to the GC cell lines. The most potent drug mestranol, an estrogen derivative, and other estrogen receptor modulators induced apoptotic events preceded by DNA damage only in the GC cell lines, but not in the GE. Moreover, mestranol could significantly suppress tumor growth of the GC cells subcutaneously transplanted into nude mice, consistent with longer survival time in the female DCKO mice than in the male. As expected, human E-cadherin-mutant and -low gastric cancer cells showed higher susceptibility to estrogen drugs in contrast to E-cadherin-intact ones in vitro and in vivo. These findings may lead to the development of novel therapeutic strategies targeting E-cadherin-deficient DGC.
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| Overall design |
The E-cadherin/p53 double conditional knockout (DCKO) mice, Atp4b-Cre+;Cdh1-loxP/loxP;Trp53-loxP/loxP genotype, were established as previously reported (Shimada et al. Gut 2012). Mice bearing tumors were sacrificed, and the primary tumors were isolated. Small pieces were immediately minced from them under sterile conditions, decolonized at 4 °C overnight in DMEM/F12 media containing 10% fetal bovine sera (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin, and subcutaneously injected into the male KSN nude mice. According to the same protocols, the transplanted tumor was dissected into aliquots which were explanted on the collagen-coated plates, and cultured in the DMEM/F12 media. The MDGC4SC1, 6 and 7 cell lines were subcloned from the MDGC4 by limiting dilution in DMEM + 10% FBS. Similarly, the MDGC7, 8 and 9 cell lines were generated from the primary cancer (MDGC7 and 8) and lymph node dissemination (MDGC9) in F12 supplemented with 5% horse or bovine sera. The GIF7, 9 and 13 cell lines have previously reported (Fukamachi et al. Biochem Biophys Res Commun 2004), and maintained in DMEM + 10% FBS.
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| Contributor(s) |
Shimada S, Akiyama Y, Mogushi K, Ishigami-Yuasa M, Kagechika H, Nagasaki H, Fukamachi H, Yuasa Y, Tanaka S |
| Citation(s) |
29527007 |
| Submission date |
Aug 07, 2017 |
| Last update date |
Jul 25, 2021 |
| Contact name |
Kaoru Mogushi |
| E-mail(s) |
mogushi-k@umin.ac.jp
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| Phone |
+81-3-5803-5184
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| Organization name |
Tokyo Medical and Dental University
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| Department |
Department of Molecular Oncology
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| Street address |
1-5-45 Yushima
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| City |
Bunkyo-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
113-8519 |
| Country |
Japan |
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| Platforms (1) |
| GPL10787 |
Agilent-028005 SurePrint G3 Mouse GE 8x60K Microarray (Probe Name version) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA397541 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE102297_RAW.tar |
95.1 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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