mRNA detection in living cells: A next generation cancer stem cell identification technique.
Cancer stem cells (CSC) are a distinct subpopulation within a tumor shown to drive tumor progression, metastasis, and recurrence. A review of the literature reveals poor consensus, with the use of a wide variety of surface markers and functional assays to identify and isolate cancer stem cells. Utilizing a novel technology that enables live-cell mRNA quantitation, we have demonstrated the ability to identify and sort viable CSC based on markers associated with stemness in pluripotent cells. Fresh tumor samples from a variety of cancer types were examined by flow cytometry for Nanog expression. Levels of CSC detected ranged from 6% to 19%. This method of CSC detection was cross-validated with other commonly used surface markers with good correlation. Matrigel invasion assays confirmed that CSC isolated using this method are both highly motile and invasive. This approach simplifies the process of identifying viable CSC from fresh tumor tissue, providing a level of accuracy not previously available. This method may also provide a valuable tool for screening and validating new CSC biomarkers.
Mitchell Cancer Institute, University of South Alabama, Mobile, AL, USA. Electronic address: smcclellan@health.southalabama.edu.
Steven
S
McClellan
Mitchell Cancer Institute, University of South Alabama, Mobile, AL, USA.
Jaroslav
J
Slamecka
Mitchell Cancer Institute, University of South Alabama, Mobile, AL, USA.
Patrick
P
Howze
Mitchell Cancer Institute, University of South Alabama, Mobile, AL, USA.
Lee
L
Thompson
Mitchell Cancer Institute, University of South Alabama, Mobile, AL, USA.
Michael
M
Finan
Mitchell Cancer Institute, University of South Alabama, Mobile, AL, USA.
Rodney
R
Rocconi
Mitchell Cancer Institute, University of South Alabama, Mobile, AL, USA.
Laurie
L
Owen
D014408:Biomarkers, Tumor / Q000032:analysis; D005434:Flow Cytometry; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000032:analysis / Q000235:genetics*; D006801:Humans; D000071317:Nanog Homeobox Protein; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis*
D014408:Biomarkers, Tumor; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D012333:RNA, Messenger
Alkaline phosphatase; Cancer stem cell; Live cell mRNA detection; Nanog; SmartFlare nanoprobe