Complete loss of miR-200 family induces EMT associated cellular senescence in gastric cancer.
Liang YuCan CaoXu LiMili ZhangQisheng GuHugh GaoJesse J BalicDuogang XuLei ZhangLe YingDakang XuYuqin YangDi WuBaokun HeBrendan J JenkinsYoudong LiuJikun LiPublished in: Oncogene (2021)
The EMT (epithelial-to-mesenchymal-transition) subtype of gastric cancer (GC) is associated with poor treatment responses and unfavorable clinical outcomes. Despite the broad physiological roles of the micro-RNA (miR)-200 family, they largely serve to maintain the overall epithelial phenotype. However, during late-stage gastric tumorigenesis, members of the miR-200 family are markedly suppressed, resulting in the transition to the mesenchymal state and the acquisition of invasive properties. As such, the miR-200 family represents a robust molecular marker of EMT, and subsequently, disease severity and prognosis. Most reports have studied the effect of single miR-200 family member knockdown. Here, we employ a multiplex CRISPR/Cas9 system to generate a complete miR-200 family knockout (FKO) to investigate their collective and summative role in regulating key cellular processes during GC pathogenesis. Genetic deletion of all miR-200s in the human GC cell lines induced potent morphological alterations, G1/S cell cycle arrest, increased senescence-associated β-galactosidase (SA-β-Gal) activity, and aberrant metabolism, collectively resembling the senescent phenotype. Coupling RNA-seq data with publicly available datasets, we revealed a clear separation of senescent and non-senescent states amongst FKO cells and control cells, respectively. Further analysis identified key senescence-associated secretory phenotype (SASP) components in FKO cells and a positive feedback loop for maintenance of the senescent state controlled by activation of TGF-β and TNF-α pathways. Finally, we showed that miR-200 FKO associated senescence in cancer epithelial cells significantly recruited stromal cells in the tumor microenvironment. Our work has identified a new role of miR-200 family members which function as an integrated unit serving to link senescence with EMT, two major conserved biological processes.
Keyphrases
- cell proliferation
- long non coding rna
- long noncoding rna
- cell cycle arrest
- endothelial cells
- rna seq
- induced apoptosis
- dna damage
- epithelial mesenchymal transition
- crispr cas
- single cell
- stress induced
- squamous cell carcinoma
- gene expression
- bone marrow
- rheumatoid arthritis
- machine learning
- transcription factor
- transforming growth factor
- high glucose
- high throughput
- high resolution
- electronic health record
- artificial intelligence
- gas chromatography
- papillary thyroid
- nucleic acid