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Phenethyl isothiocyanate suppresses stemness in the chemo- and radio-resistant triple-negative breast cancer cell line MDA-MB-231/IR via downregulation of metadherin

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Author(s)
Yen Thi Kim Nguyen
Issued Date
2020
URI
http://dcoll.jejunu.ac.kr/common/orgView/000000009354
Abstract
Resistance to chemotherapy and radiation therapy is considered a major therapeutic barrier in breast cancer. Cancer stem cells (CSCs) play a prominent role in chemo- and radiotherapy resistance. The established chemo- and radio-resistant triple-negative breast cancer (TNBC) cell line MDA-MB-231/IR displays greater CSC characteristics than the parental MDA-MB-231 cells. Escalating evidence demonstrates that metadherin (MTDH) is associated with a number of cancer signaling pathways as well as breast cancer therapy resistance, making it an attractive therapeutic target. Kaplan–Meier plot analysis revealed a correlation between higher levels of MTDH and shorter lifetimes in breast cancer and TNBC patients. Moreover, there was a positive correlation between the MTDH and CD44 expression levels in The Cancer Genome Atlas breast cancer database. We demonstrate that MTDH plays a pivotal role in the regulation of stemness in MDA-MB-231/IR cells. Knockdown of MTDH in MDA-MB-231/IR cells resulted in a reduction in the CSC population, aldehyde dehydrogenase activity, and major CSC markers, including β-catenin, CD44+, and Slug. In addition, MTDH knockdown increased reactive oxygen species (ROS) levels in MDA-MB-231/IR cells. We found that phenethyl isothiocyanate, a well-known pro oxidant phytochemical, suppressed stemness in MDA-MB-231/IR cells through ROS modulation via downregulation of MTDH.
Awarded Date
2020. 2
Table Of Contents
ABSTRACT 1
1. INTRODUCTION . 2
2. MATERIALS AND METHODS 5
2.1. Cell culture . 5
2.2. Cell viability assay 5
2.3. Cell migration assay . 5
2.4. Cell invasion assay 6
2.5. Flow cytometric analysis of the CD44+/CD24 population 6
2.6. ALDEFLUOR assay 6
2.7. Mammosphere formation assay 7
2.8. Real-time PCR 7
2.9. Western blot analysis 7
2.10. siRNA knockdown 8
2.11. ROS generation analysis 8
2.12. GSH assay 9
2.13. Correlation analysis by the KaplanMeier Plotter . 9
2.14. Statistical analysis . 9
3. RESULTS 10
3.1. Stem cell characteristics of MDA-MB-231/IR cells . 10
3.2. MDA-MB-231/IR cells exhibited low ROS levels 12
3.3. MTDH expression is significantly correlated with poorer prognosis in breast cancer patient samples . 14
3.4. MTDH plays a key role in maintenance of the CSC population in MDA-MB-231/IR cells . 16
3.5. PEITC reduced MDA-MB-231/IR cell viability by inducing ROS accumulation . 18
3.6. PEITC reduced stemness and the expression of MTDH and CSC markers in MDA MB-231/IR cells . 20
3.7. PEITC downregulated MTDH at the post-transcriptional level 24
4. DISCUSSION 26
5. CONCLUSIONS . 30
REFERENCES . 32
Degree
Master
Publisher
제주대학교 대학원
Citation
Yen Thi Kim Nguyen. (2020). Phenethyl isothiocyanate suppresses stemness in the chemo- and radio-resistant triple-negative breast cancer cell line MDA-MB-231/IR via downregulation of metadherin
Type
Dissertation
Appears in Collections:
General Graduate School > Interdisciplinary Graduate Program in Advanced Convergence Technology and Science
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