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Published on: July 22, 2020
Identification of downstream metastasis-associated target genes regulated by LSD1 in colon cancer cells
Jiang Chen1, Jie Ding2, Ziwei Wang1
1Department of Gastrointestinal Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Purpose:
This study aims to identify downstream target genes regulated by lysine-specific demethylase 1 (LSD1) in colon cancer cells and investigate the molecular mechanisms of LSD1 influencing invasion and metastasis of colon cancer.
Method:
We obtained the expression changes of downstream target genes regulated by small-interfering RNA-LSD1 and LSD1-overexpression via gene expression profiling in two human colon cancer cell lines. An Affymetrix Human Transcriptome Array 2.0 was used to identify differentially expressed genes (DEGs). We screened out LSD1-target gene associated with proliferation, metastasis, and invasion from DEGs via Gene Ontology and Pathway Studio. Subsequently, four key genes (CABYR, FOXF2, TLE4, and CDH1) were computationally predicted as metastasis-related LSD1-target genes. ChIp-PCR was applied after RT-PCR and Western blot validations to detect the occupancy of LSD1-target gene promoter-bound LSD1.
Result:
A total of 3633 DEGs were significantly upregulated, and 4642 DEGs were downregulated in LSD1-silenced SW620 cells. A total of 4047 DEGs and 4240 DEGs were upregulated and downregulated in LSD1-overexpressed HT-29 cells, respectively. RT-PCR and Western blot validated the microarray analysis results. ChIP assay results demonstrated that LSD1 might be negative regulators for target genes CABYR and CDH1. The expression level of LSD1 is negatively correlated with mono- and dimethylation of histone H3 lysine4(H3K4) at LSD1- target gene promoter region. No significant mono-methylation and dimethylation of H3 lysine9 methylation was detected at the promoter region of CABYR and CDH1.
Conclusion:
LSD1- depletion contributed to the upregulation of CABYR and CDH1 through enhancing the dimethylation of H3K4 at the LSD1-target genes promoter. LSD1- overexpression mediated the downregulation of CABYR and CDH1expression through decreasing the mono- and dimethylation of H3K4 at LSD1-target gene promoter in colon cancer cells. CABYR and CDH1 might be potential LSD1-target genes in colon carcinogenesis.
Insights
Lysine-specific demethylase 1 (LSD1) regulates colon cancer metastasis by controlling CABYR and CDH1 gene expression. LSD1 depletion upregulates these genes via H3K4 methylation, while LSD1 overexpression downregulates them.
Area of Science:
- Molecular Biology
- Oncology
- Epigenetics
Background:
- Lysine-specific demethylase 1 (LSD1) is implicated in cancer progression.
- Understanding LSD1's role in colon cancer metastasis is crucial for therapeutic development.
Purpose of the Study:
- Identify downstream target genes of LSD1 in colon cancer cells.
- Elucidate the molecular mechanisms by which LSD1 influences colon cancer invasion and metastasis.
Main Methods:
- Gene expression profiling (microarray) was used to identify differentially expressed genes (DEGs) after LSD1 manipulation (siRNA and overexpression) in colon cancer cell lines.
- Bioinformatic analysis (Gene Ontology, Pathway Studio) screened DEGs for roles in proliferation, metastasis, and invasion.
- Quantitative PCR, Western blot, and ChIP assay validated target genes (CABYR, FOXF2, TLE4, CDH1) and LSD1's regulatory mechanism.
Main Results:
- Thousands of DEGs were identified upon LSD1 silencing or overexpression.
- LSD1 was confirmed as a negative regulator of CABYR and CDH1 expression.
- LSD1's activity correlated with H3K4 methylation levels at target gene promoters.
Conclusions:
- LSD1 depletion upregulates CABYR and CDH1 by enhancing H3K4 dimethylation at their promoters.
- LSD1 overexpression downregulates CABYR and CDH1 by decreasing H3K4 methylation.
- CABYR and CDH1 are potential LSD1 target genes in colon carcinogenesis.
