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| Status |
Public on Oct 02, 2020 |
| Title |
DNA methylation analysis of cloned HCT116 cells treated with a DNA demethylating drug. |
| Organism |
Homo sapiens |
| Experiment type |
Methylation profiling by genome tiling array
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| Summary |
Epigenetic reprogramming using demethylating drugs is a promising approach for cancer therapy, but its efficacy is highly dependent on the dosing regimen. Low-dose treatment for a prolonged period shows a high therapeutic efficacy, despite its small demethylating effect. Here, we aimed to reveal the mechanisms of how such low-dose treatment shows high efficacy by focusing on epigenetic reprograming at the single-cell level. Single-cell RNA-sequencing of HCT116 cells treated with decitabine (DAC) revealed that up-regulated genes were highly variable at the single-cell level. To analyze functional consequences at the single-cell level, DAC-treated HCT116 cells were cloned. While only partial reduction of methylation levels was observed in bulk cells, complete demethylation of specific cancer-related genes was observed, depending upon clones. For example, p16 was completely demethylated in the H3-32 clone out of 9 clones, and this clone showed slower proliferation than other clones without demethylation. In addition, in this clone, the fraction of cells with tetraploid became much larger, indicating that cellular senescence was induced. These results showed that epigenetic reprogramming of specific cancer-related pathways at the single-cell level is likely to underlie the high efficacy of low-dose DNA demethylating therapy.
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| Overall design |
DNA methylation statuses of cloned HCT116 cells were analyzed using Infinium HumanMethylation450.
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| Contributor(s) |
Takeshima H, Yoda Y, Watanabe N, Ushijima T |
| Citation(s) |
32958049 |
| |
| Submission date |
Apr 23, 2020 |
| Last update date |
Apr 07, 2021 |
| Contact name |
Hideyuki Takeshima |
| E-mail(s) |
takeshima.hideyuki@hoshi.ac.jp
|
| Organization name |
Hoshi University
|
| Department |
Institute for Advanced Life Sciences
|
| Lab |
Department of Epigenomics
|
| Street address |
2-4-41 Ebara
|
| City |
Shinagawa-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
142-8501 |
| Country |
Japan |
| |
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| Platforms (1) |
| GPL13534 |
Illumina HumanMethylation450 BeadChip (HumanMethylation450_15017482) |
|
| Samples (11)
|
| GSM4494968 |
DNA methylation in mock-treated bulk HCT116 cells |
| GSM4494969 |
DNA methylation in decitabine-treated bulk HCT116 cells |
| GSM4494970 |
DNA methylation in clone H3-01 |
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| This SubSeries is part of SuperSeries: |
| GSE171622 |
Low-dose DNA demethylating therapy induces reprogramming of diverse cancer-related pathways at the single-cell level |
|
| Relations |
| BioProject |
PRJNA627908 |
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