|ZFIN ID: ZDB-PUB-191022-11|
CRISPR-Cas9 Knockdown and Induced Expression of CD133 Reveal Essential Roles in Melanoma Invasion and Metastasis
Simbulan-Rosenthal, C.M., Dougherty, R., Vakili, S., Ferraro, A.M., Kuo, L.W., Alobaidi, R., Aljehane, L., Gaur, A., Sykora, P., Glasgow, E., Agarwal, S., Rosenthal, D.S.
|Source:||Cancers 11(10): (Journal)|
|Registered Authors:||Glasgow, Eric|
|Keywords:||CRISPR-Cas9 knockdown, invasion, matrix metalloproteinases, melanoma initiating cells, metastasis|
|PubMed:||31623313 Full text @ Cancers|
Simbulan-Rosenthal, C.M., Dougherty, R., Vakili, S., Ferraro, A.M., Kuo, L.W., Alobaidi, R., Aljehane, L., Gaur, A., Sykora, P., Glasgow, E., Agarwal, S., Rosenthal, D.S. (2019) CRISPR-Cas9 Knockdown and Induced Expression of CD133 Reveal Essential Roles in Melanoma Invasion and Metastasis. Cancers. 11(10):.
ABSTRACTCD133, known as prominin1, is a penta-span transmembrane glycoprotein presumably a cancer stem cell marker for carcinomas, glioblastomas, and melanomas. We showed that CD133(+) 'melanoma-initiating cells' are associated with chemoresistance, contributing to poor patient outcome. The current study investigates the role(s) of CD133 in invasion and metastasis. Magnetic-activated cell sorting of a melanoma cell line (BAKP) followed by transwell invasion assays revealed that CD133(+) cells are significantly more invasive than CD133(-) cells. Conditional reprogramming of BAKP CD133(+) cells maintained stable CD133 overexpression (BAK-R), and induced cancer stem cell markers, melanosphere formation, and chemoresistance to kinase inhibitors. BAK-R cells showed upregulated CD133 expression, and consequently were more invasive and metastatic than BAK-P cells in transwell and zebrafish assays. CD133 knockdown by siRNA or CRISPR-Cas9 (BAK-R-T3) in BAK-R cells reduced invasion and levels of matrix metalloproteinases MMP2/MMP9. BAK-R-SC cells, but not BAK-R-T3, were metastatic in zebrafish. While CD133 knockdown by siRNA or CRISPR-Cas9 in BAK-P cells attenuated invasion and diminished MMP2/MMP9 levels, doxycycline-induced CD133 expression in BAK-P cells enhanced invasion and MMP2/MMP9 concentrations. CD133 may therefore play an essential role in invasion and metastasis via upregulation of MMP2/MMP9, leading to tumor progression, and represents an attractive target for intervention in melanoma.
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