Crosstalk-eliminated Quantitative Determination of Aflatoxin B1-induced Hepatocellular Cancer Stem Cells Based on Concurrent Monitoring of CD133, CD44, and Aldehyde Dehydrogenase1
CD133, CD44, ALDH1의 하이콘텐트 모니터링에 기초한 아플라톡신 B1 유발 간암 줄기세포 정량
- Hee Ju
- 약학대학 약학과
- Issue Date
- 서울대학교 대학원
- Hepatocellular cancer stem cell; Aflatoxin B1; High-content cellular imaging; Quantum dot; Diagnosis markers
- 학위논문 (석사)-- 서울대학교 대학원 : 약학과, 2016. 8. 송준명.
- Cancer stem cells (CSCs), known as tumor initiating cells, have become a critically important issue for cancer therapy. The properties of CSCs, such as anticancer drug and radiation resistance, have been considered to be the source of postoperative recurrence and metastasis. 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). Although much research has demonstrated the induction of hepatocellular cancer by aflatoxin B1 found in contaminated foods and the interrelation between them, the formation of hepatocellular CSCs caused by aflatoxin B1 and their quantitative determination are hardly reported. Hepatocelluar CSCs were acquired from HepG2 cells 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 aflatoxin B1. The results matched well with those obtained utilizing flow cytometry, which is used as a general tool for the detection of hepatocellular CSCs. This study demonstrates that accurate quantitative measurement of aflatoxin B1-induced hepatocellular CSCs can be accomplished using a constructed multicolor cellular imaging system that eliminates crosstalk among hepatocellular CSC biomarkers.