PIK3CA-altered head and neck cancer- specific antitumor effect and underlying mechanism of nonsteroidal anti- inflammatory drugs.
- 주제(키워드) Head and neck cancer , PIK3CA mutation , NSAIDs , ER stress , Reactive Oxygen Species , Mitochondrial dysfunction.
- 주제(DDC) 570
- 발행기관 아주대학교 일반대학원
- 지도교수 Lee Sangyoon
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000033415
- 본문언어 한국어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Certain nonsteroidal anti-inflammatory drugs (NSAIDs) are well-known as not only Cyclooxygenase-2 (COX-2) inhibitors but also anti-tumor drugs for many types of cancer worldwide, including head and neck squamous cell carcinoma (HNSCC). It is claimed that mutated PIK3CA oncogenes are found in several human solid tumors. The oncogenic properties of PIK3CA, in particular, contribute to the development of human HNSCC. Some research shows that the anti-tumor effect of NSAIDs is more effective in PIK3CA-alternated HNSCCs, which highly express COX-2. However, the mechanisms have not been studied yet. In this study, we aimed to investigate some underlying mechanisms of Celecoxib and Sulindac Sulfide in PIK3CA-mutated HNSSC. FaDu and Detroit562 cell lines was used for wild-type and mutated PIK3CA models of HNSCCs, respectively. These two NSAIDs against tumor growth of the Detroit562 cell line by provoking endoplasmic reticulum (ER) stress and reactive oxygen species (ROS). Simultaneously, both Celecoxib and Sulindac Sulfide promote the expression of Death Receptor 5 (DR5), cleaved-caspase 3, and some mitochondria-related cell death markers such as BAX and BIM, and mitochondrial damage in altered-PIK3CA cells, resulting in significantly declined cell growth compared to the wild-type. Therefore, this study provides useful information to improve the efficiency of chemotherapy for PIK3CA-altered HNSCC patients. Keywords: Head and neck cancer; PIK3CA mutation; NSAIDs; ER stress; Reactive Oxygen Species; Mitochondrial dysfunction.
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I. INTRODUCTION 1
A. Head and neck squamous cell carcinoma and current treatment approaches 1
B. The Phosphoinositide 3-kinase (PI3K) structure and activity. 4
C. Characteristic of PIK3CA mutation and its function in cancer development. 8
D. NSAIDs are also used as anti-cancer drugs in HNSCC. 10
E. The aim of study. 12
II. MATERIALS AND METHODS 14
1. Chemicals and antibodies . 14
2. Cell culture and chemical treatments 14
3. Cell viability assay . 15
4. Western blot analysis 15
5. Immunofluorescence 16
6. Quantitative Reverse transcription-PCR (qRT-PCR) 16
7. COX-2 gene knockdown 17
8. Intracellular ROS and superoxide analysis 17
9. Measurement of mitochondrial membrane potential . 18
10. PGE2 enzyme-linked immunosorbent assay (Elisa) . 18
11. Colony Formation Assays . 18
12. Spheroid formation assay . 19
13. Statistical Analysis. 19
III. RESULTS 20
1. The huge difference in COX-2 expression between PI3K wild-type and mutant HNSCCs. 20
2. NSAIDs significantly suppressed the cell viability of mt-PIK3CA HNSCCs. 24
3. PIK3CA mutation promotes the NSAID's efficiency in inducing ER stress, resulting in the elevation of DR5 expression 27
4. Several NSAIDs induce intracellular ROS accumulation 33
5. Crosstalk between ER stress and ROS generated by Celecoxib and SUS-treated PIK3CA-mutated HNSCC cells 35
6. Celecoxib and SUS treatment results in mitochondrial dysfunction and nuclear localization of cleaved caspase-3 in PIK3CA-mutated HNC cells 38
7. ER stress and ROS production are linked to celecoxib-induced mitochondrial malfunction, which results in PIK3CA-mutated HNC cell death 40
IV. DISCUSSION 42
V. CONCLUSION 47
VI. REFERENCES 48
