CDK4/6 inhibitors induce senescence in the breast cancer cell lines with enhanced anti- tumor immunogenic properties compared with DNA-damaging agents
- 주제(키워드) Therapy-induced senescence , CDK4/6 inhibitors , DNA-damaging agents , Senescence-associated secretory phenotype , Tumor microenvironment
- 주제(DDC) 570
- 발행기관 아주대학교 일반대학원
- 지도교수 Tae Jun Park
- 발행년도 2024
- 학위수여년월 2024. 8
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000034144
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
CDK4/6 inhibitors induce senescence in the breast cancer cell lines with enhanced anti-tumor immunogenic properties compared with DNA- damaging agents Understanding the phenomenon of senescence during cancer treatment remains controversial, yet crucial for comprehending its impact on tumor suppression or activation. Therefore, this study aims to compare and analyze the senescence status of breast cancer cells induced by various chemotherapeutic agents (DNA-damaging agents and CDK4/6 inhibitors). Although these agents induce a similar extent of senescence, it has been identified and observed that senescence-associated secretory phenotype (SASP), which promotes inflammatory cytokines (TGFβ, TNFA and IL-6), pro-tumorigenic immunity (CXCL2, CXCL8 and CSF1), and pro-angiogenic factors (VEGF family and GDF15), are more robust in senescent tumor cells induced by DNA-damaging agents. Interestingly, senescent tumor cells induced by cyclin- dependent kinase4/6 inhibitors (CDK4/6i) not only exhibit increased expression of SASP and ligands associated with anti-tumor immunity (HLA family and B2M) but also release chemokines (CXCL9, 10, and 11) into the tumor microenvironment (TME) to enhance immune surveillance and recruit NK and T cells compared to DNA- damaging agents. Despite the deficient activation of Nuclear Factor-Kappa-B (NF-κB) and p53 pathway in CDK4/6i-induced senescence, SASP expressions are still comparable to those of DNA-damaging agents-induced senescence. Overall, this study shows that tumor cells treated with various types of agents elicit senescence to similar extents, while the expressions of pro-tumorigenic and anti-tumorigenic SASP vary substantially between the agents, depending on the type of chemotherapeutic agents used. Thus, TIS by CDK4/6i may serve as an effective approach for treatment strategies in cancer patients by utilizing the distinct characteristics of senescence. Keywords: Therapy-induced senescence, CDK4/6 inhibitors, DNA-damaging agents, Senescence-associated secretory phenotype, Tumor microenvironment
more목차
I. INTRODUCTUON 1
1. Cellular senescence 1
2. Cancer senescence 4
3. Therapy-induced senescence 6
4. Remodeling of the TME by TIS 10
5. Proposal 13
II. MATERIALS AND METHOD 14
1. Cell culture 14
2. Reagents for senescence in the breast cancer cell lines 14
3. Induction of senescence in the breast cancer cell lines 14
4. Assessment of SA-β-galactosidase activity 15
5. Fluorescence Activated Cell Sorting (FACS) 15
6. Cell viability assay 15
7. RNA Extraction and quantitative real-time PCR 15
8. Western blotting 16
9. Enzyme-Linked Immunosorbent Assay (ELISA) 16
10. Tube formation assay 17
11. RNA sequencing and normalization of differentially expressed genes (DEGs) 17
12. Analysis of transcriptome and data visualization 17
13. Data availability statement 18
14. Statistical analysis 18
15. List of primer sequences 19
III. RESULTS 21
1. DNA-damaging agents and CDK4/6i induce a comparable level of senescence in breast cancer cells 21
2. Aromatase inhibitors (AI) exhibit no impact on CDK4/6i-induced senescence 26
3. TIS by DNA-damaging agents and CDK4/6i is distinctively regulated by genes associated with inflammation, tumor immunity, and angiogenesis 29
4. DNA-damaging agents-induced senescence demonstrate elevated expression levels of pro-tumorigenic cytokines and ligands compared to CDK4/6i 38
5. Angiogenesis is more abundantly enhanced by DNA-damaging agents- than by CDK4/6i-induced senescent tumor cells 46
6. The activities of antigen presentation and interferon signaling in CDK4/6i-induced senescence tumor cells are comparably enhanced with those of DNA-damaging agents 55
7. DNA-damaging agents and CDK4/6i induce senescence in HCC1428 breast cancer cells similar to MCF-7 64
8. TIS by DNA-damaging agents (Etoposide and Carboplatin) or CDK4/6i (Abemaciclib) differently regulate inflammatory cytokines and angiogenesis regulators, immune-related factors in breast cancer cell line 71
9. The activation of p53 and NF-κB signaling is notably more pronounced in senescence induced by DNA-damaging agents compared to CDK4/6i 76
IV. DISCUSSION 83
1. Senescence induced by various therapeutic agents 83
2. Pro-tumorigenic effects of TIS 84
3. Angiogenesis regulation of TIS 85
4. Anti-tumor immune activation of TIS 87
5. Signaling pathway of TIS 88
6. Implications and limitations 89
7. Further study 90
Ⅴ. CONCLUSION 92
Ⅵ. REFERENCE 94
국문요약 125
