Table 2.
Mechanisms associated with cancer therapy-related cardiovascular aging.
| Treatment modalities | Experimental subjects | Treatment protocols | Detection of senescence | Mechanisms | Year |
|---|---|---|---|---|---|
| Anthracycline Drugs | In vitro: Isolated neonatal rat cardiomyocytes | In vitro: Treatment with 10−7 mol/L DOX for 7 days | Increased SA-β-gal activity; Decreased in telomerase activity; Shortened telomere length; Increased protein levels of P27kip1 and p21cip1/waf1; Increased mRNA levels of p16INK4a | The activation of the PML-acetylated p53 complex | 2008 (47) |
| In vivo: 4-week-old male Wistar rats | In vivo: Intraperitoneal administration of DOX, divided into six equal injections (2.5 mg/kg each) over 2 weeks, with a total cumulative dose of 15 mg/kg body weight, with sample collection for monitoring at 11 months of age | ||||
| In vitro: Isolated Ventricular Myocytes from 2-Day-Old Sprague-Dawley Rats (Lonza) and H9c2 Rat Cardiomyoblasts | In vitro: Treatment with 0.1 μM DOX for 3 h, followed by assessment at 24 and 48 h | Increased SA-β-gal activity; Cellular senescence morphology; Increased micronuclei; Chromosomal abnormalities; Increased protein levels of p53 | The downregulation of TRF1 and TRF2 via MAPK and p53-mediated pathways | 2009 (48) | |
| In vitro: Isolated c-kit positive human cardiac progenitor cells | In vitro: Treatment with 0.1, 0.5, and 1.0 μM DOX for 24 and 48 h, followed by 7 days of culture | Increased SA-β-gal activity; Increased tissue immunofluorescence labeling of p16INK4a and γ-H2AX; Decreased expression of BrdU and Ki67; Activation of DNA damage pathways | The activation of the p16-Rb pathway-triggered cell cycle arrest | 2013 (30) | |
| In vivo: Human cardiac autopsy specimens | In vivo: Standard clinical doses for cancer patients | ||||
| In vitro: Human primary umbilical artery vascular smooth muscle cells (VSMCs) and mouse aortic VSMCs. | In vitro: Treatment with 0.25, 0.5, and 1 µM DOX for 3 h, followed by replacement with normal culture medium and monitoring after 3 days | Increased SA-β-Gal activity; Decreased cell viability; Increased protein levels of p53, p21Cip1/Waf1, and p16INK4a | The upregulation of uPAR-triggered TRF2 ubiquitination and proteasomal degradation | 2013 (49) | |
| In vitro: Human vascular smooth muscle cells | In vitro: Treatment of passages 5–8 cells with 100 nM DOX to induce premature senescence, followed by analysis at 1, 3, and 7 days | Increased SA-β-gal activity; Cellular senescence morphology; Cell cycle arrest; Increased DNA damage-associated 53BP1 foci; Increased micronuclei formation; Increased γ-H2AX levels; Increased protein levels of Increased p21cip1/waf1 protein levels; Increased expression of SASP; Increased superoxide production; DNA methylation inhibition | The Activation of DNA Damage Repair-related ATM Pathway-coupled with Superoxide Level Accumulation | 2014 (50) | |
| In vitro: H9c2 cells and isolated primary cardiomyocytes from mice. | In vitro: Pre-treatment with 0.1 μmol/L DOX for 3 h, followed by sample collection after 21 h, 45 h, 7 days, 14 days, and 21 days in normal culture medium | Increased SA-β-gal activity; Increased protein levels of p53, and p16INK4a | The downregulation of TRF2-induced reduction in telomere protection | 2016 (51) | |
| In vivo: 4-week-old male Wistar rats | In vivo: Subcutaneous injection of DOX at 2 mg/kg once weekly for 7 weeks, total cumulative dose of 14 mg/kg, with sample collection in week 9 | Reduced mitochondrial DNA (mtDNA); Decreased DNA methylation levels | The depletion of mtDNA-coupled with DNA methylation reduction | 2017 (52) | |
| In vitro: HL-1 mouse cardiomyocytes | In vitro: Treatment with 5 µM DOX for 0, 24, 48, and 72 h | Decreased cell viability; Shortened telomere length; Decreased in telomerase activity; Elevated intracellular ROS; Elevated mitochondrial superoxide; Increased mRNA levels of p53, p27Kip1, and p16INK4a | The increase in lincRNA-p21 expression-coupled with Wnt/β-catenin Signaling Pathway Activation and ROS-related Indicator Increase | 2018 (53) | |
| In vitro: H9c2 cells | In vitro: Treatment with 0.5 µM DOX for 72 h | Decreased cell viability; Increased mRNA levels of p53 and p16INK4a; Shortened telomere length; Decreased in telomerase activity | The Elevation of TGF-β1 Expression | 2018 (54) | |
| In vitro: H9c2 rat embryonic cardiomyocytes | In vitro: Treatment with 0.1 µM DOX for 24 h | Increased SA-β-gal activity | The miR-34a/PNUTS axis | 2019 (55) | |
| In vitro: H9c2 cells | In vitro: Treatment with 0.1 µM DOX for 48 h | Increased SA-β-gal activity; Decreased cell viability; Increased protein levels of p21cip1/waf1 and p16INK4a; Increased expression of SASP; Elevated intracellular ROS | The upregulation of TXNIP-triggering intracellular redox imbalance and NLRP3 inflammasome activation | 2020 (56) | |
|
In vitro: Isolated neonatal rat cardiomyocytes and H9c2 cells; In vivo: 3–4 week old Wistar rats |
In vitro: Treatment with 50 nM DOX for 3 h, followed by monitoring after 7 days; In vivo: Treatment with DOX through intraperitoneal injections 9 times every other day at 5 mg/kg, with a total cumulative dose of 45 mg/kg, collect samples after 8 months |
Increased SA-β-gal activity; Reduced mtDNA; Increased numbers of p16INK4a positive cells and myosin positive cells under flow cytometry. | The damage of mtDNA | 2020 (57) | |
| In vitro: Cardiomyocytes differentiated from human induced pluripotent stem cells (iPSC) | In vitro: Treatment with 0.5 μM DOX for 24 h | Increased SA-β-gal activity; Cell cycle arrest; Increased mRNA levels of p53 and p21cip1/waf1 | The upregulation of miR-92a-3p and inhibition of ATG4a expression-induced attenuation of mitochondrial metabolism | 2020 (58) | |
|
In vitro: Isolated mouse ventricular myocytes; In vivo: eight-week-old male C57BL/6 mice |
In vitro: Treatment with 1 μM DOX for 72 h; In vivo: Intraperitoneal injection of DOX was carried out on Mondays, Wednesdays and Fridays within one week, at a dose of 4 mg/kg each time, with a total cumulative dose of 12 mg/kg. Sample collection for monitoring was initiated on the 14th day |
Increased SA-β-gal activity; Increased mRNA levels of p27kip1, p16INK4a, and p21cip1/waf1 | The activation of the miR-221–3p/Sirt2 pathway | 2020 (59) | |
| In vitro: Isolated neonatal mouse cardiomyocytes | In vitro: Treatment with 1 μM DOX for 24 h and 7 days | Increased SA-β-gal activity; Increased γ-H2AX foci; Decreased cell viability; Increased protein levels of p53 and p21cip1/waf1; Elevated mitochondrial ROS; Elevated intracellular ROS; Increased expression of SASP | The downregulation of mitochondrial autophagy via TBK1 K63-linked polyubiquitination facilitation | 2021 (60) | |
| In vitro: H9c2 cells | In vitro: Treatment with 0.5 µM DOX for 24 h, followed by replacement with normal culture medium and monitoring after 10 days | Increased SA-β-gal activity; Decreased cell viability; Shortened telomere length; Increased intracellular ROS and mitochondrial superoxide; Increased mRNA levels of p53 and p21cip1/waf1 | The depletion of Sirt6-caused mitochondrial damage, telomere dysfunction, increased H3K9 acetylation and upregulated NF-κB-related oxidative stress | 2021 (61) | |
|
In vitro: H9c2 cells; In vivo: C57BL/6 mice |
In vitro: Pre-treatment with 0.1 µM DOX for 3 h, followed by replacement with normal culture medium and monitoring after 0, 24, 48 and 72 h; In vivo: Intraperitoneal injections at 2.5 mg/kg, three times a week for two consecutive weeks, with a cumulative dose of 15 mg/kg, collect samples for monitoring 4 months after completion of injections |
Increased SA-β-gal activity; Decreased cell viability; Increased protein levels of p53, p21Cip1/Waf1, and p16INK4a; Increased expression of SASP | The p38 MAPK-Redd1-NF-κB pathway | 2021 (62) | |
| In vitro: H9c2 rat embryonic cardiomyocytes and AC16 human cardiomyocyte-like cells | In vitro: Treatment with 0.1 μmol/l DOX for 24 h | Increased SA-β-gal activity; Shortened telomere length; Decreased in telomerase activity; Increased protein levels of p53, p21cip1/waf1, p16INK4a, and IGFBP3 | The upregulation of C5a and C5aR-induced elevation of TNF-α and IFN-γ expression and ROS level | 2021 (63) | |
|
In vivo: Human ventricular cardiac fibroblasts (HCF), human umbilical vein endothelial cells (HUVECs); In vitro: 6-week-old wild-type BALB/c female mice |
In vivo: Treatment with 100 nM DOX for 7 days, followed by replacement with serum-free medium and monitoring after 2 days; In vitro: Inject dox dissolved in 0.9% saline subcutaneously every other day starting on the 7th day after tumor formation, with a cumulative dose of 22 mg/kg, collect samples after 3 weeks |
Increased protein levels of p21cip1/waf1 and p16INK4a | / | 2022 (64) | |
| In vitro: H9c2 cells | In vitro: Pre-treatment with 0.3 μM DOX for 24 h | Increased SA-β-gal activity; Decreased cell viability; Shortened telomere length; Increased intracellular ROS and mitochondrial superoxide; Increased mRNA levels of p53 and p21cip1/waf1 | The Klotho/SIRT1 signaling pathway. | 2022 (65) | |
| In vitro: Immortalized human umbilical vein endothelial cell line EA.hy926 and primary HUVECs | In vitro: Treatment with 0.5µM DOX for 24 h, followed by replacement with normal culture medium and monitoring after 72 h and 5 days | Increased SA-β-gal activity; Cell cycle arrest; Increased protein levels of p53 and p21cip1/waf1; Increased expression of SASP; | The promotion of anti-apoptotic protein expression | 2022 (66) | |
|
In vitro: Cardiomyocytes differentiated from iPSC (iCM) and isolated mouse cardiomyocytes; In vivo: Cardiac tissue transcriptomics data |
In vitro: Treatment with sub-lethal concentration of 0.2 μM DOX for 3 h, followed by replacement with normal culture medium and monitoring after 4 days | Increased SA-β-gal activity; Cell cycle arrest; Increased γ-H2AX positive nuclear foci; Increased mRNA and protein levels of p21cip1/waf1 and p16INK4a; Increased expression of SASP; Increased cell size; Elevated intracellular ROS | The loss of mitochondrial membrane potential-coupled with ROS increase | 2022 (67) | |
| In vitro: HL-1 cardiomyocytes |
In vitro: Treatment with 100 nM DOX for 72 h; In vivo: Administer a total cumulative dose of 20 mg/kg intraperitoneally in 8 injections over 4 weeks at 2.5 mg/kg per injection, and collect samples on the 44th day |
Increased SA-β-gal activity; Cell cycle arrest; Decreased Ki67 proliferation; Increased mRNA and protein levels of p53 and p21cip1/waf1 | The promotion of anti-apoptotic protein expression | 2022 (68) | |
| In vivo: 10-week-old male and female C57BL/6 J mice | |||||
| In vitro: HUVECs | In vitro: Treatment with 100 nM DOX for 24 h. | Increased SA-β-gal activity; Decreased cell proliferation; Increased protein levels of p21cip1/waf1 and p16INK4a | The ALDH1A2/AKT/ERK1/2-p21 pathway | 2022 (69) | |
| In vitro: HUVECs and EA.hy926 human endothelial-derived cell line | In vitro: Treatment with 0.5 μM DOX for 24 h, followed by replacement with normal culture medium and monitoring after 72 and 120 h | Increased SA-β-gal activity; Increased mRNA levels of p53 and p21cip1/waf1 | The activation of MAPK and JNK pathways-coupled with NF-κB-related oxidative stress and SASP | 2023 (70) | |
|
In vitro: H9c2 cells; In vivo: 8-week-old C57BL/6 female mice |
In vitro: Treatment with 0.1 μM DOX for 6 days; In vivo: Intraperitoneal injections at 3 mg/kg, once daily for 7 days, with a cumulative dose of 21 mg/kg, collect samples for monitoring on the 9th day |
Increased SA-β-gal activity; Decreased cell viability; Increased mRNA levels of p53 and p16INK4a; Increased expression of SASP; Elevated intracellular ROS | The promotion of mTOR protein phosphorylation | 2023 (71) | |
|
In vitro: Isolated neonatal mouse cardiomyocytes (NMCM); In vivo: 6–8-week-old ICR mice |
In vitro: Treatment with 1 μM DOX for 72 h; In vivo: Treat ICR mice with intraperitoneal injections of DOX at 3 mg/kg, 6 times within two weeks, with a total cumulative dose of 18 mg/kg, collect samples on the 35th day |
Increased SA-β-gal activity; Increased mRNA and protein levels of p21cip1/waf1 and p16INK4a; Increased mitochondrial fragmentation | The activation of the VPO1/ERK pathway-triggering enhanced mitochondrial fission | 2024 (72) | |
|
In vitro: AC16 cells; In vivo: C57BL/6 mice |
In vitro: Treatment with 1.25 μM, 2.5 μM, and 5 μM DOX for 24 h; In vivo: Intraperitoneal injections at 2.5 mg/kg, three times a week for two consecutive weeks, with a cumulative dose of 15 mg/kg, collect samples for monitoring 4 months after completion of injections |
Increased SA-β-gal activity; Decreased cell viability; Increased mRNA and protein levels of p27kip1, p16INK4a, and p21cip1/waf1; Increased expression of SASP | The suppression of CRIF1 expression and promotion of PXDN expression-facilitating mitochondrial fission and oxidative stress | 2024 (73) | |
|
In vitro: H9c2 cells; In vivo: 8-week-old male C57/Bl6 mice |
In vitro: Pre-treatment with 1 μM DOX for 24 h; In vivo: Single intraperitoneal injection of 20 mg/kg DOX, collect samples for monitoring after 2 weeks |
Increased SA-β-gal activity; Increase γ-H2AX foci; Increased mRNA levels of IGFBP3, p21cip1/waf1 and p16INK4a; Increased expression of SASP; Elevated intracellular ROS | The upregulation of PARP-2 expression, suppression of SIRT1 expression and activity, and activation of the FOXO1/p53 signaling pathway | 2024 (74) | |
|
In vitro: H9c2 cells; In vivo: C57BL/6 mice |
In vitro: Treatment with 1.0 μM DOX for 0, 12, 24, and 48 h; In vivo: single intraperitoneal injection of 20 mg/kg, collect samples for monitoring after two weeks |
Increased SA-β-gal activity; Increase γ-H2AX foci; Decreased cell viability; Increased mRNA levels of p53, p21cip1/waf1 and p16INK4a; Increased expression of SASP; Increased superoxide dismutase (SOD) activity; Elevated intracellular ROS | The depletion of SIRT6 and downregulation of PPARα | 2024 (75) | |
| In vivo: C57BL/6 mice | In vivo: Administer a total cumulative dose of 24 mg/kg intraperitoneally in 6 injections over 6 weeks at 4 mg/kg per injection, and collect samples after 4 days | Increased mRNA levels of p53, p21cip1/waf1, p16INK4a and p19Arf; Increased expression of SASP; | The activation of the p38/MAPK signaling pathway | 2024 (76) | |
| In vitro: Human cardiac organoids (hCOs or hCardioids) | In vitro: Treatment with 0.5 μM DOX for 3 and 32 days; | Decreased expression of Ki67; Increased mRNA levels of p15, p21cip1/waf1, p16INK4a and p19Arf; Elevated intracellular ROS | The activation of the oxidative stress pathway | 2025 (77) | |
|
In vitro: Cardiomyocytes differentiated from human induced pluripotent stem cells (iPSC); In vivo: C57BL/6 mice |
In vitro: Treatment with 1.0 μM DOX for 24 h; In vivo: Administer a total cumulative dose of 20 mg/kg intraperitoneally in 4 injections over 4 weeks at 5 mg/kg per injection, and collect samples at the end of the 6th week |
Increased SA-β-gal activity; Increased mRNA and protein levels of p53, p16INK4a, and p21cip1/waf1; Elevated mitochondrial ROS | The leakage of mtRNA and the activation of the cGAS-STING pathway | 2025 (78) | |
| Radiotherapy | In vitro: Bovine Aortic Endothelial Cells (BAECs) and HUVECs | In vitro: Sample collection 3–5 days after 8 Gy ionizing radiation | Increased SA-β-gal activity; Cellular senescence morphology; Cell cycle arrest; Increased γ-H2AX levels; Increased expression of SASP | The damage of DNA | 2007 (79) |
| In vitro: HL-1 mouse cardiomyocytes and H9c2 rat cardiomyocytes | In vitro: Sample collection at 72 h and 96 h post-exposure to 0, 2, and 8 Gy ionizing radiation | Increased SA-β-gal activity; Decreased cell proliferation; Cellular senescence morphology; Elevated intracellular ROS | The accumulation of ROS | 2015 (80) | |
| In vitro: Human aortic endothelial cells | In vitro: Sample collection at 4 and 6 days post-irradiation with unknown Gy dose | Increased SA-β-gal activity; Decreased cell proliferation; Cellular senescence morphology; Increased γ-H2AX levels; Increased protein levels of p53, p21Cip1/Waf1, and p16INK4a | The increase of GDF15 expression, ROS accumulation and activation of ERK signaling pathway along with p16/Rb pathway | 2016 (81) | |
| In vitro: Human cardiomyocytes (HCMs) | In vitro: Sample collection at 24, 48, and 72 h post-exposure to 5 Gy ionizing radiation | Increased SA-β-gal activity; Decreased cell proliferation; Increased mRNA levels of p21Cip1/Waf1 and p16INK4a; Elevated intracellular ROS | The inhibition of SIRT1, upregulation of miR-34a expression and induction of oxidative stress | 2018 (82) | |
| In vivo: Carotid arteries of apolipoprotein E knockout (ApoE−/−) mice | In vivo: After 2 weeks of inducing atherosclerotic lesions by ligating the left carotid artery, perform 6Gy whole-body irradiation on mice, collect samples after 4 weeks | Increase DNA damage-related 53BP1 foci; Increased γ-H2AX levels; Increased protein levels of p16INK4a; Increased mRNA levels of p21Cip1/Waf1 and p16INK4a | The DNA damage-coupled with SASP and other inflammatory factors | 2021 (83) |