| Title |
A novel method to generate single-cell-derived cancer-associated fibroblast clones
|
|---|---|
| Published in |
Journal of Cancer Research and Clinical Oncology, March 2017
|
| DOI | 10.1007/s00432-017-2409-3 |
| Pubmed ID | |
| Authors |
Hiroko Hashimoto, Yoshitaka Suda, Tomoyuki Miyashita, Atsushi Ochiai, Masahiro Tsuboi, Kenkichi Masutomi, Tohru Kiyono, Genichiro Ishii |
| Abstract |
Cancer-associated fibroblasts (CAFs) communicate with cancer cells to play important roles in tumor progression. However, CAFs have heterogeneous phenotypes and functions. To understand how much of this heterogeneity relates to different biological responses, a more efficient method of generating single-cell-derived CAF clones is required. We transduced two primary CAF cultures (CAFs-608 and CAFs-621) from lung adenocarcinoma with human telomerase reverse transcriptase (hTERT), mutant forms of cyclin dependent kinase 4 (CDK4R24C) independently and in combination (hTERT/CDK4R24C). After live imaging of each sorted-single cell, we evaluated the numbers of successfully established clones from CAFs-hTERT, CAFs-CDK4R24C, and CAFs-hTERT/CDK4R24C. Furthermore, we examined the expression levels of genes associated with tumor promoting pathways in established clones by qRT-PCR. Overexpression of hTERT and CDK4R24C efficiently extended the lifespan of both CAFs-608 and CAFs-621. The number of established CAF clones was highest for CAFs-hTERT/CDK4R24C, with 57 and 62 clones established from CAFs-608 and CAFs-621, respectively. Conversely, 16 and 11 CAFs-hTERT clones were derived from CAFs-608 and CAFs-621, respectively and 10 and 8 CAFs-CDK4R24C clones were from CAFs-608 and CAFs-621, respectively. TGF-b, ATCA2, and HSF1 mRNA levels differed in individual clones established from CAFs-hTERT/CDK4R24C. The expression levels of ATCA2 and HSF1 were much higher in one clone than in the other established clones and the parental CAFs. Our results show that combined exogenous expression of hTERT and mutant CDK4 is an effective method to generate single-cell-derived CAF clones. This provides an innovative and suitable approach to investigate the heterogeneous function and phenotype of CAFs. |
X Demographics
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Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 24 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 24 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 9 | 38% |
| Student > Bachelor | 3 | 13% |
| Other | 2 | 8% |
| Student > Doctoral Student | 2 | 8% |
| Student > Master | 2 | 8% |
| Other | 2 | 8% |
| Unknown | 4 | 17% |
| Readers by discipline | Count | As % |
|---|---|---|
| Medicine and Dentistry | 8 | 33% |
| Biochemistry, Genetics and Molecular Biology | 7 | 29% |
| Agricultural and Biological Sciences | 2 | 8% |
| Immunology and Microbiology | 2 | 8% |
| Neuroscience | 1 | 4% |
| Other | 0 | 0% |
| Unknown | 4 | 17% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 25 July 2018.
All research outputs
#19,221,261
of 23,815,455 outputs
Outputs from Journal of Cancer Research and Clinical Oncology
#1,814
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#237,477
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Outputs of similar age from Journal of Cancer Research and Clinical Oncology
#21
of 41 outputs
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