Anticancer effects of Adefovir dipivoxil and its action mechanisms in two human breast cancer cell lines
- 주제(키워드) Adefovir dipivoxil , breast cancer , replication stress , reactive oxygen species , apoptosis
- 주제(DDC) 570
- 발행기관 아주대학교 일반대학원
- 지도교수 Jong-Soo Lee
- 발행년도 2024
- 학위수여년월 2024. 8
- 학위명 석사
- 학과 및 전공 일반대학원 생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000033842
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Despite a variety of therapeutic strategies for breast cancers treatment, the survival rate of breast cancer patients has not been improved due to its increasing resistance to radio- and chemo-therapy. Since drug repurposing for breast cancer treatment can lead to significant cost and time savings compared to new drug development, I have investigated the anticancer effects and action mechanisms of Adefovir dipivoxil (ADV), an FDA-approved antiviral nucleoside analogue used in the therapy of human hepatitis B virus, in two human breast cancer cell lines: MCF7 (wild-type p53, estrogen receptor (ER), and progesterone receptor (PR)-positive) and MDA-MB-231 (mutated p53 and triple hormone receptors-negative). ADV exhibited anti-proliferative effects on both breast cancer cell lines, suggesting that ADV may have anticancer effects in a p53- and hormone receptors-independent manner. ADV induced DNA replication stress, leading to stalling of cell cycle progression at S-phase and activation of surrogate DNA damage signaling ATM-CHK2 and ATR-CHK1 pathways in both cell lines. Interestingly, the apoptosis hallmarks, cleavages of caspases and poly(ADP-ribose) polymerase (PARP-1) were observed in both ADV-treated MCF7 and MDA-MB-231 cells, but DNA laddering and Annexin V-positive cell number was detected only in ADV-treated MCF7 cells, suggesting that ADV-mediated cell death pathways in two cell lines might be differential. In addition, reactive oxygen species (ROS) level was highly enhanced in MDA-MB-231 cells but it was marginally increased in MCF7 cells by ADV. The increased ROS-activated mitogen-activated protein kinases (MAPKs), p38, JNK1/2, and ERK1/2 could lead to necrosis, suggesting that ADV promotes apoptotic and necrotic induction in MDA-MB-231 cells. In conclusion, I herein demonstrate the potential of ADV as a novel therapeutic agent for breast cancer treatment including triple-negative breast cancer, which tends to grow and spread quickly but have, has fewer treatment options. Keywords: Adefovir dipivoxil, breast cancer, replication stress, reactive oxygen species, apoptosis
more목차
I. INTRODUCTION 2
II. MATERIALS AND METHODS . 4
1. Cells and Reagents . 4
2. Cell Proliferation Assay 4
3. Cell Cycle Analysis. 4
4. Clonogenic assay . 5
5. BrdU Incorporation Assay 5
6. Western Blot Analysis 6
7. Apoptosis Assays . 6
8. Cytosolic DNA fragmentation . 7
9. Small interfering RNAs (siRNAs) 7
10. Statistical analysis 7
III. RESULTS 8
1. ADV suppressed cellular proliferation of human breast cancer cell lines . 8
2. ADV induced replication stress and S-phase arrest in breast cancer cells . 10
3. ADV-induced replication stress activated DNA damage signaling in breast cancer cells. 14
4. ADV induced apoptosis in breast cancer cells. 16
5. UCN-01 synergistically increased anti-proliferative effects of ADV in MCF7 cells. 22
6. UCN-01 increased ADV-induced apoptosis in MCF7 cells . 25
7. Apoptosis induction in MCF7 cells is CHK2-p53-dependent . 28
8. ADV induced ROS generation and MAPK-dependent apoptosis in MDA-MB-231 cell. 33
IV. DISCUSSION 38
REFERENCE 43
