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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Nov 30, 2019 |
| Title |
PROX1 mediates chemoresistance-associated recurrence via maintenance of quiescent colon cancer stem cells |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Cancer stem cells (CSCs) are profoundly associated with refractory nature of cancer. A quiescent population of CSCs is responsible for tumorigenesis and chemoresistance in leukemia, whereas neither the presence nor clinical importance of the quiescent CSCs is clearly established in solid tumors. In colon cancer, LGR5 is regarded as a functional marker of CSCs, but heterogeneity among LGR5+ cells was not clearly defined. Here we stratified LGR5+ cells by single-cell gene expression analyses and revealed that a tumorigenic population of mouse LGR5+ cells resides in a quiescent state. We identified 23 signature genes that were uniquely expressed in the quiescent CSCs of mouse tumors, and found that 7 of them were also specifically expressed in a quiescent population of LGR5+ cells in human colon xenograft tumors. Among them, PROX1 was expressed in invasive fronts of colon tumors. PROX1 was induced by TCF1 (TCF7), and the TCF1-mediated PROX1 expression was responsible for not only the maintenance of quiescence but also chemoresistance of colon cancer organoids. Knockout of PROX1 in patient-derived xenograft tumors resulted in inhibition of tumor recurrence after chemotherapeutic treatment. Our data underscore the therapeutic importance of a quiescent CSC population in colon cancer.
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| Overall design |
Three individual tumors were obtained from a DSS-administrated ApcMin/+ mouse and used in this study. The tumors were mixed after the enzymatic digestion and DAPI-Cd31-Cd45-Epcam+Cd24+ cells were single-cell sorted into a 96-well plate containing lysis buffer [0.5 % (v/v) NonidetP-40 and 2 U/µl SUPERase-In (Ambion)] and stored frozen at -80 °C. The frozen cells were lysed and subjected for cDNA synthesis using Bead-seq method described in previous reports (DOI:10.1016/j.ab.2014.10.01, DOI:10.1038/s41598-017-04616-6). The processes from the production of cDNA libraries to their amplification and first purification were carried out automatically using a Caliper Zephyr compact liquid handler (Perkin Elmer, CA, USA) coupled with a homemade operation program. The cDNA library produced from each cell was used to examine Lgr5 expression by semi-quantitative PCR reaction for (32 cycles) using KOD polymerase (Takara), followed by Ethidium Bromide (Sigma) staining. Lgr5-positive cDNAs were then converted in to the pooled libraries for Illumina sequencing using the Nextera XT DNA preparation kit (Illumina).
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| Contributor(s) |
Shiokawa D, Sakai H, Ohata H, Miyazaki T, Kanda Y, Sekine S, Hosokawa M, Narushima D, Kato M, Suzuki Y, Takeyama H, Kambara H, Nakagama H, Okamoto K |
| Citation(s) |
32816913 |
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| Submission date |
Nov 29, 2019 |
| Last update date |
Aug 23, 2020 |
| Contact name |
Daichi Narushima |
| Organization name |
National Cancer Center Research Institute
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| Department |
Department of Bioinformatics
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| Street address |
5-1-1 Tsukiji, Chuo-ku
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| City |
Tokyo |
| ZIP/Postal code |
104-0045 |
| Country |
Japan |
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| Platforms (1) |
| GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
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| Samples (56)
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| Relations |
| BioProject |
PRJNA592447 |
| SRA |
SRP234017 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE141157_tpm_gene_id.csv.gz |
1.2 Mb |
(ftp)(http) |
CSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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