CRISPR-Cas9 Knockdown and Induced Expression of CD133 Reveal Essential Roles in Melanoma Invasion and Metastasis
| Autor: | Anirudh Gaur, Dean S. Rosenthal, Eric Glasgow, Leala Aljehane, Ryan Dougherty, Alexandra M Ferraro, Cynthia M. Simbulan-Rosenthal, Li-Wei Kuo, Sahar Vakili, Peter Sykora, Ryyan Alobaidi, Seema Agarwal |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Cancer Research
MMP2 crispr-cas9 knockdown Biology lcsh:RC254-282 Article Metastasis 03 medical and health sciences fluids and secretions 0302 clinical medicine Cancer stem cell medicine metastasis neoplasms 030304 developmental biology 0303 health sciences Gene knockdown Melanoma matrix metalloproteinases Cell sorting medicine.disease invasion lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens 3. Good health carbohydrates (lipids) melanoma initiating cells Oncology Tumor progression 030220 oncology & carcinogenesis embryonic structures cardiovascular system Cancer research biological phenomena cell phenomena and immunity Reprogramming |
| Zdroj: | Cancers, Vol 11, Iss 10, p 1490 (2019) Cancers Volume 11 Issue 10 |
| ISSN: | 2072-6694 |
| Popis: | CD133, known as prominin1, is a penta-span transmembrane glycoprotein presumably acancer stem cell marker for carcinomas, glioblastomas, and melanomas. We showed that CD133(+)&lsquo melanoma-initiating cells&rsquo are associated with chemoresistance, contributing to poor patientoutcome. The current study investigates the role(s) of CD133 in invasion and metastasis. Magneticactivatedcell sorting of a melanoma cell line (BAKP) followed by transwell invasion assays revealedthat CD133(+) cells are significantly more invasive than CD133(&minus ) cells. Conditional reprogrammingof BAKP CD133(+) cells maintained stable CD133 overexpression (BAK-R), and induced cancer stemcell markers, melanosphere formation, and chemoresistance to kinase inhibitors. BAK-R cellsshowed upregulated CD133 expression, and consequently were more invasive and metastatic thanBAK-P cells in transwell and zebrafish assays. CD133 knockdown by siRNA or CRISPR-Cas9 (BAKR-T3) in BAK-R cells reduced invasion and levels of matrix metalloproteinases MMP2/MMP9. BAKR-SC cells, but not BAK-R-T3, were metastatic in zebrafish. While CD133 knockdown by siRNA orCRISPR-Cas9 in BAK-P cells attenuated invasion and diminished MMP2/MMP9 levels,doxycycline-induced CD133 expression in BAK-P cells enhanced invasion and MMP2/MMP9concentrations. CD133 may therefore play an essential role in invasion and metastasis viaupregulation of MMP2/MMP9, leading to tumor progression, and represents an attractive target forintervention in melanoma. |
| Databáze: | OpenAIRE |
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