CRISPR/Cas9-mediated knockout of PIM3 suppresses tumorigenesis and cancer cell stemness in human hepatoblastoma cells
| Autor: | Raoud Marayati, Juliet L. Easlick, Laura L. Stafman, David K. Crossman, Laura V. Bownes, Elizabeth Mroczek-Musulman, Elizabeth A. Beierle, Jerry E. Stewart, Hooper R. Markert, Colin H. Quinn, Adele P. Williams |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Hepatoblastoma Cancer Research Carcinogenesis Cell Biology Protein Serine-Threonine Kinases medicine.disease_cause 03 medical and health sciences Mice 0302 clinical medicine Downregulation and upregulation Cell Line Tumor Proto-Oncogene Proteins medicine Animals Humans Molecular Biology Cell Proliferation Kinase Liver Neoplasms medicine.disease digestive system diseases 030104 developmental biology medicine.anatomical_structure Cell Transformation Neoplastic Cell culture Apoptosis 030220 oncology & carcinogenesis Cancer cell Cancer research Molecular Medicine CRISPR-Cas Systems |
| Zdroj: | Cancer gene therapy. 29(5) |
| ISSN: | 1476-5500 |
| Popis: | Hepatoblastoma remains one of the most difficult childhood tumors to treat and is alarmingly understudied. We previously demonstrated that Proviral Insertion site in Maloney murine leukemia virus (PIM) kinases, specifically PIM3, are overexpressed in human hepatoblastoma cells and function to promote tumorigenesis. We aimed to use CRISPR/Cas9 gene editing with dual gRNAs to introduce large inactivating deletions in the PIM3 gene and achieve stable PIM3 knockout in the human hepatoblastoma cell line, HuH6. PIM3 knockout of hepatoblastoma cells led to significantly decreased proliferation, viability, and motility, inhibited cell-cycle progression, decreased tumor growth in a xenograft murine model, and increased animal survival. Analysis of RNA sequencing data revealed that PIM3 knockout downregulated expression of pro-migratory and pro-invasive genes and upregulated expression of genes involved in apoptosis and differentiation. Furthermore, PIM3 knockout decreased hepatoblastoma cancer cell stemness as evidenced by decreased tumorsphere formation, decreased mRNA abundance of stemness markers, and decreased cell surface expression of CD133, a marker of hepatoblastoma stem cell-like cancer cells. Reintroduction of PIM3 into PIM3 knockout cells rescued the malignant phenotype. Successful CRISPR/Cas9 knockout of PIM3 kinase in human hepatoblastoma cells confirmed the role of PIM3 in promoting hepatoblastoma tumorigenesis and cancer cell stemness. |
| Databáze: | OpenAIRE |
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