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Baicalein suppresses stem cell-like characteristics of resistant human breast cancer cells MDA-MB-231 by up-regulation of interferon-induced protein with tetratricopeptide repeats 2

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Author(s)
Koh, So Yae
Issued Date
2019
URI
http://dcoll.jejunu.ac.kr/common/orgView/000000009180
Abstract
Breast cancer is the second leading cause of cancer death for woman and the most prevalent type of cancer. Triple negative breast cancer (TNBC) is the most poor prognosis subtype of breast cancer and represented with high tumor grade and metastasis. We generated radio- and chemo- resistance cells by irradiation (IR), MDA-MB-231/IR. The resistance cells were enhanced in migration (1.72-fold), invasion (1.48-fold), and stem cell￾like characteristics such as mammosphere formation (1.44-fold) and side population (1.67- fold). Pathway analysis exhibited that MDA-MB-231/IR had activated including NF-κB pathway (11%), TNF signaling pathway (10%), and Toll-like receptor pathway (10%). Within 77 differentially expressed genes (DEGs) demonstrated by our transcriptome analysis, 12 DEGs were associated with cancer resistance on MDA-MB-231/IR; the genes were AKR1C1 (6.83log2 fold), FTL (6.64-log2 fold), MK (2.92-log2 fold), LG3BP (6.01-log2 fold), GFPT2 (3.04-log2 fold), TGFBI (3.34-log2 fold), AKR1C3 (3.87-log2 fold), CCDC69 (2.96-log2 fold), AKR1C2 (3.87-log2 fold), TWF1, PKP3, and IFIT2 (-2.84-log2 fold). Baicalein induced apoptosis on MDA-MB-231/IR and the half maximum inhibitory concentration (IC 50) of baicalein was 39.58 ± 2.86 μM. Baicalein reversed radio- and chemo- resistance, migration (0.61-fold), invasion (0.43-fold), mammosphere formation (0.12-fold), and expression of CD44high and CD24low population of MDA-MB-231/IR (0.48- fold) while increasing the expression of interferon-induced protein with tetratricopeptide repeats 2 (IFIT2) (3-fold in mRNA and 27-fold in protein level). Using the cancer genome atlas (TCGA) database, IFIT2 expression was concerned with metastasis and poor prognosis marker for TNBC patients.
Affiliation
제주대학교 대학원
Department
대학원 차세대융복합과학기술협동과정
Advisor
김소미
Awarded Date
2019. 8
Table Of Contents
1. Abstract. 1
2. Introduction 2
2.1. Breast cancer
2.2. Cancer stem cells
2.3. Epithelial to mesenchymal transition
2.4. Invasion and metastasis
2.5. Phytochemicals
2.6. Baicalein
2.7. Interferon-induced proteins with tetratricopeptide repeat
2.8. Transcriptomic analysis
2.9. Summary 7
3. Materials and methods 9
3.1. Reagents
3.2. Cell culture and resistant cell generation
3.3. Cell viability
3.4. Cell morphology
3.5. Clonogenic assay
3.6. Mammosphere
3.7. RNA extraction, cDNA synthesis, and Realtime PCR
3.8. Wound healing assay
3.9. Invasion assay
3.10. Flow cytometry
3.11. Western blot
3.12. Transcriptomic analysis
3.13. Pathway analysis
3.14. Gene expression analysis
3.15. Statistical analyses
4. Results. 16
4.1. MDA-MB-231/IR cells developed radio-resistance
4.2. MDA-MB-231/IR cells developed chemo-resistance
4.3. Chemo- resistance of MDA-MB-231/IR cells evaluated by Annexin V/PI assay
4.4. MDA-MB-231/IR cells exhibited epithelial to mesenchymal morphology compared to parental cells
4.5. MDA-MB-231/IR cells exhibited enhanced migration and invasion ability compared toparental cells
4.6. Mammosphere forming ability and expression levels of stem cell-like markers were enhanced in MDA-MB-231/IR cells than those of parental cells
4.7. MDA-MB-231/IR cells exhibited stem cell-like characters compared to its parental cells
4.8. Transcriptomics analysis on MDA-MB-231/IR cells
4.9. Cancer resistance was reported in 12 DEGs of MDA-MB-231/IR
4.10. Baicalein effectively targeted chemo- and radio-resistance MDA-MB-231/IR cells
4.11. Baicalein reversed expression of IFIT2 in MDA-MB-231/IR
4.12. Baicalein reduced protein expression level of Slug
4.13. Baicalein inhibited EMT capacity of MDA-MB-231/IR
4.14. Baicalein diminished the mammosphere formation and expression of marker proteins in MDA-MB-231/IR
4.15. Baicalein diminished the stem cell-like characters of MDA-MB-231/IR
4.16. Baicalein increased expression levels of apoptosis-related proteins on MDAMB-231/IR
4.17. Baicalein induces apotosis on MDA-MB-231/IR cells
4.18. Baicalein co-treatment reversed radio- resistance in MDA-MB-231/IR
4.19. Baicalein co-treatment reversed chemo- resistance in MDA-MB-231/IR
4.20. The expression of IFIT2 affects survival in patients
5. Discussion. 59
6. References. 70
Degree
Doctor
Publisher
제주대학교 대학원
Citation
Koh, So Yae. (2019). Baicalein suppresses stem cell-like characteristics of resistant human breast cancer cells MDA-MB-231 by up-regulation of interferon-induced protein with tetratricopeptide repeats 2
Type
Dissertation
Appears in Collections:
General Graduate School > Interdisciplinary Graduate Program in Advanced Convergence Technology and Science
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