title: Crosstalk-eliminated quantitative determination of aflatoxin B1-induced hepatocellular cancer stem cells based on concurrent monitoring of CD133, CD44, and aldehyde dehydrogenase1.
abstract: Cancer stem cells (CSCs), known as tumor initiating cells, have become a critically important issue for cancer therapy. Although much research has demonstrated the induction of hepato cellular carcinoma by aflatoxin B1, the formation of hepatocellular CSCs and their quantitative determination is hardly reported. In this work, it was found that hepatocellular CSCs were produced from HepG2 cells by aflatoxin B1-induced mutation, and their amount was quantitatively determined using crosstalk-eliminated multicolor cellular imaging based on quantum dot (Qdot) nanoprobes and an acousto-optical tunable filter (AOTF). Hepatocellular CSCs were acquired via magnetic bead-based sorting and observed using concurrent detection of three different markers: CD133, CD44, and aldehyde dehydrogenase1 (ALDH1). The DNA mutation of HepG2 cells caused by aflatoxin B1 was quantitatively observed via absorbance spectra of aflatoxin B1-8, 9-epoxide-DNA adducts. The percentages of hepatocellular CSCs formed in the entire HepG2 cells were determined to be 9.77±0.65%, 10.9±1.39%, 11.4±1.32%, and 12.8±0.7%, respectively, at 0 μM, 5 μM, 10 μM, and 20 μM of aflatoxin B1. The results matched well with those obtained utilizing flow cytometry. This study demonstrates that aflatoxin mediated mutation induced the conversion of hepatic cancer cell to hepatic CSCs by using a Qdot based constructed multicolor cellular imaging system.
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Hee
authors.initials: H
authors.lastname: Ju
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Shim
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Parthasarathy
authors.initials: P
authors.lastname: Arumugam
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 151-742, South Korea. Electronic address: jmsong@snu.ac.kr.
authors.forename: Joon Myong
authors.initials: JM
authors.lastname: Song
mesh_terms: D000071916:AC133 Antigen; D016604:Aflatoxin B1 / Q000633:toxicity*; D000444:Aldehyde Dehydrogenase / Q000378:metabolism*; D015703:Antigens, CD / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000139:chemically induced / Q000378:metabolism; D018736:DNA Adducts / Q000187:drug effects*; D006023:Glycoproteins / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D008113:Liver Neoplasms / Q000139:chemically induced / Q000378:metabolism; D009154:Mutation; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000473:pathology; D010455:Peptides / Q000378:metabolism*
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; C497483:CD44 protein, human; D018736:DNA Adducts; D006023:Glycoproteins; D018960:Hyaluronan Receptors; C000606023:PROM1 protein, human; D010455:Peptides; D016604:Aflatoxin B1; D000444:Aldehyde Dehydrogenase
keywords: Aflatoxin B(1); Diagnosis markers; Hepatocellular cancer stem cell; High-content cellular imaging; Quantum dot