Mesenchymal stem cells confer resistance to doxorubicin‑induced cardiac senescence by inhibiting microRNA‑34a

Xia,Wenzheng; Hou,Meng

doi:10.3892/ol.2018.8438

Mesenchymal stem cells confer resistance to doxorubicin‑induced cardiac senescence by inhibiting microRNA‑34a

Authors:
- Wenzheng Xia
- Meng Hou
View Affiliations

Affiliations: Department of Neurosurgery, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China, Department of Radiation Oncology, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China
Published online on: April 23, 2018 https://doi.org/10.3892/ol.2018.8438
Pages: 10037-10046

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Doxorubicin (DOXO) is a chemotherapeutic agent widely used in the treatment of various types of cancer. However, cardiotoxicity is a major side effect of DOXO therapy due to the ability of this compound to induce cardiac cellular senescence. It is well known that microRNA (miR)‑34a serves a role in cardiac dysfunction and ageing, and that it is involved in several cellular processes associated with DOXO‑induced cardiotoxicity. Furthermore, mesenchymal stem cell (MSC)‑based therapies have been reported to modulate cellular senescence. In the present study, the Transwell system was used to co‑culture H9c2 cells and MSCs, and cell proliferation and viability were assessed. The expression of senescence‑related genes, p53 and p16, and telomere length were analyzed using reverse transcription‑quantitative polymerase chain reaction (PCR), and the protein expression levels of situin 1 (SIRT1) were detected by western blotting. Additionally, telomerase activity of H9c2 was examined using the Telo TAGGG Telomerase PCR ELISA PLUS kit. The present study revealed that, in the presence of DOXO, H9c2 cells were in senescence, as characterized by a low proliferation rate, poor viability and a marked increase in the expression of p53 and p16. By contrast, when co‑cultured with MSCs in the presence of DOXO, the proliferation and viability of H9c2 cells increased. Additionally, the expression of p53 and p16 decreased, and increased length of telomere and telomerase activity was also observed. Additionally, the mechanism underlying the anti‑senescence function of MSCs was revealed to involve the miR‑34a‑SIRT1 axis, confirmed by overexpressing miR‑34a using a miR‑34a mimic or silencing SIRT1 using small interfering RNA, which abolished the anti‑senescence effect of MSCs on DOXO‑treated H9c2 cells. Taken together, the results of the present study suggest that MSCs may rejuvenate H9c2 cells from a state of DOXO‑induced senescence by increasing SIRT1 expression, there by inhibiting miR‑34a. Therefore, treatment with MSCs may have important therapeutic implications in the restoration of cardiotoxicity in patients with cancer undergoing treatment with DOXO.

View Figures

View References

1	Rochette L, Guenancia C, Gudjoncik A, Hachet O, Zeller M, Cottin Y and Vergely C: Anthracyclines/trastuzumab: New aspects of cardiotoxicity and molecular mechanisms. Trends Pharmacol Sci. 36:326–348. 2015. View Article : Google Scholar : PubMed/NCBI
2	Stone JB and DeAngelis LM: Cancer-treatment-induced neurotoxicity-focus on newer treatments. Nat Rev Clin Oncol. 13:92–105. 2016. View Article : Google Scholar : PubMed/NCBI
3	Suter TM and Ewer MS: Cancer drugs and the heart: Importance and management. Eur Heart J. 34:1102–1111. 2013. View Article : Google Scholar : PubMed/NCBI
4	Franco VI, Henkel JM, Miller TL and Lipshultz SE: Cardiovascular effects in childhood cancer survivors treated with anthracyclines. Cardiol Res Pract. 2011:1346792011. View Article : Google Scholar : PubMed/NCBI
5	Volkova M and Russell R III: Anthracycline cardiotoxicity: Prevalence, pathogenesis and treatment. Curr Cardiol Rev. 7:214–220. 2011. View Article : Google Scholar : PubMed/NCBI
6	Ezquer F, Gutiérrez J, Ezquer M, Caglevic C, Salgado HC and Calligaris SD: Mesenchymal stem cell therapy for doxorubicin cardiomyopathy: Hopes and fears. Stem Cell Res Ther. 6:1162015. View Article : Google Scholar : PubMed/NCBI
7	Yu Q, Li Q, Na R, Li X, Liu B, Meng L, Liutong H, Fang W, Zhu N and Zheng X: Impact of repeated intravenous bone marrow mesenchymal stem cells infusion on myocardial collagen network remodeling in a rat model of doxorubicin-induced dilated cardiomyopathy. Mol Cell Biochem. 387:279–285. 2014. View Article : Google Scholar : PubMed/NCBI
8	Garbade J, Dhein S, Lipinski C, Aupperle H, Arsalan M, Borger MA, Barten MJ, Lehmann S, Walther T and Mohr FW: Bone marrow-derived stem cells attenuate impaired contractility and enhance capillary density in a rabbit model of Doxorubicin-induced failing hearts. J Card Surg. 24:591–599. 2009. View Article : Google Scholar : PubMed/NCBI
9	Eulalio A, Mano M, Dal Ferro M, Zentilin L, Sinagra G, Zacchigna S and Giacca M: Functional screening identifies miRNAs inducing cardiac regeneration. Nature. 492:376–381. 2012. View Article : Google Scholar : PubMed/NCBI
10	Seeger FH, Zeiher AM and Dimmeler S: MicroRNAs in stem cell function and regenerative therapy of the heart. Arterioscler Thromb Vasc Biol. 33:1739–1746. 2013. View Article : Google Scholar : PubMed/NCBI
11	Yang Y, Cheng HW, Qiu Y, Dupee D, Noonan M, Lin YD, Fisch S, Unno K, Sereti KI and Liao R: MicroRNA-34a plays a key role in cardiac repair and regeneration following myocardial infarction. Circ Res. 117:450–459. 2015. View Article : Google Scholar : PubMed/NCBI
12	Boon RA, Iekushi K, Lechner S, Seeger T, Fischer A, Heydt S, Kaluza D, Tréguer K, Carmona G, Bonauer A, et al: MicroRNA-34a regulates cardiac ageing and function. Nature. 495:107–110. 2013. View Article : Google Scholar : PubMed/NCBI
13	Chen F and Hu SJ: Effect of microRNA-34a in cell cycle, differentiation, and apoptosis: A review. J Biochem Mol Toxicol. 26:79–86. 2012. View Article : Google Scholar : PubMed/NCBI
14	Piegari E, Russo R, Cappetta D, Esposito G, Urbanek K, Dell'Aversana C, Altucci L, Berrino L, Rossi F and De Angelis A: MicroRNA-34a regulates doxorubicin-induced cardiotoxicity in rat. Oncotarget. 7:62312–62326. 2016. View Article : Google Scholar : PubMed/NCBI
15	Haigis MC and Sinclair DA: Mammalian sirtuins: Biological insights and disease relevance. Annu Rev Pathol. 5:253–295. 2010. View Article : Google Scholar : PubMed/NCBI
16	Morris BJ: Seven sirtuins for seven deadly diseases of aging. Free Radic Biol Med. 56:133–171. 2013. View Article : Google Scholar : PubMed/NCBI
17	Deng CX: SIRT1, is it a tumor promoter or tumor suppressor? Int J Biol Sci. 5:147–152. 2009. View Article : Google Scholar : PubMed/NCBI
18	Luo J, Nikolaev AY, Imai S, Chen D, Su F, Shiloh A, Guarente L and Gu W: Negative control of p53 by Sir2alpha promotes cell survival under stress. Cell. 107:137–148. 2001. View Article : Google Scholar : PubMed/NCBI
19	Yang G, Zhang X, Weng X, Liang P, Dai X, Zeng S, Xu H, Huan H, Fang M, Li Y, et al: SUV39H1 mediated SIRT1 trans-repression contributes to cardiac ischemia-reperfusion injury. Basic Res Cardiol. 112:222017. View Article : Google Scholar : PubMed/NCBI
20	De Angelis A, Piegari E, Cappetta D, Russo R, Esposito G, Ciuffreda LP, Ferraiolo FA, Frati C, Fagnoni F, Berrino L, et al: SIRT1 activation rescues doxorubicin-induced loss of functional competence of human cardiac progenitor cells. Int J Cardiol. 189:30–44. 2015. View Article : Google Scholar : PubMed/NCBI
21	Liu WH, Liu JJ, Wu J, Zhang LL, Liu F, Yin L, Zhang MM and Yu B: Novel mechanism of inhibition of dendritic cells maturation by mesenchymal stem cells via interleukin-10 and the JAK1/STAT3 signaling pathway. PLoS One. 8:e554872013. View Article : Google Scholar : PubMed/NCBI
22	Xia W, Zhang F, Xie C, Jiang M and Hou M: Macrophage migration inhibitory factor confers resistance to senescence through CD74-dependent AMPK-FOXO3a signaling in mesenchymal stem cells. Stem Cell Res Ther. 6:822015. View Article : Google Scholar : PubMed/NCBI
23	Xia W, Xie C, Jiang M and Hou M: Improved survival of mesenchymal stem cells by macrophage migration inhibitory factor. Mol Cell Biochem. 404:11–24. 2015. View Article : Google Scholar : PubMed/NCBI
24	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
25	Crepin T, Carron C, Roubiou C, Gaugler B, Gaiffe E, Simula-Faivre D, Ferrand C, Tiberghien P, Chalopin JM, Moulin B, et al: ATG-induced accelerated immune senescence: Clinical implications in renal transplant recipients. Am J Transplant. 15:1028–1038. 2015. View Article : Google Scholar : PubMed/NCBI
26	Zhang F, Cui J, Liu X, Lv B, Liu X, Xie Z and Yu B: Roles of microRNA-34a targeting SIRT1 in mesenchymal stem cells. Stem Cell Res Ther. 6:1952015. View Article : Google Scholar : PubMed/NCBI
27	Bao Y, Yin M, Hu X, Zhuang X, Sun Y, Guo Y, Tan S and Zhang Z: A safe, simple and efficient doxorubicin prodrug hybrid micelle for overcoming tumor multidrug resistance and targeting delivery. J Control Release. 235:182–194. 2016. View Article : Google Scholar : PubMed/NCBI
28	Tanaka M, Koul D, Davies MA, Liebert M, Steck PA and Grossman HB: MMAC1/PTEN inhibits cell growth and induces chemosensitivity to doxorubicin in human bladder cancer cells. Oncogene. 19:5406–5412. 2000. View Article : Google Scholar : PubMed/NCBI
29	Liu J, Mao W, Ding B and Liang CS: ERKs/p53 signal transduction pathway is involved in doxorubicin-induced apoptosis in H9c2 cells and cardiomyocytes. Am J Physiol Heart Circ Physiol. 295:H1956–H1965. 2008. View Article : Google Scholar : PubMed/NCBI
30	Burridge PW, Li YF, Matsa E, Wu H, Ong SG, Sharma A, Holmström A, Chang AC, Coronado MJ, Ebert AD, et al: Human induced pluripotent stem cell-derived cardiomyocytes recapitulate the predilection of breast cancer patients to doxorubicin-induced cardiotoxicity. Nat Med. 22:547–556. 2016. View Article : Google Scholar : PubMed/NCBI
31	Golpanian S, Wolf A, Hatzistergos KE and Hare JM: Rebuilding the damaged heart: Mesenchymal stem cells, cell-based therapy, and engineered heart tissue. Physiol Rev. 96:1127–1168. 2016. View Article : Google Scholar : PubMed/NCBI
32	Samper E, Diez-Juan A, Montero JA and Sepúlveda P: Cardiac cell therapy: Boosting mesenchymal stem cells effects. Stem Cell Rev. 9:266–280. 2013. View Article : Google Scholar : PubMed/NCBI
33	Zhao SL, Zhang YJ, Li MH, Zhang XL and Chen SL: Mesenchymal stem cells with overexpression of midkine enhance cell survival and attenuate cardiac dysfunction in a rat model of myocardial infarction. Stem Cell Res Ther. 5:372014. View Article : Google Scholar : PubMed/NCBI
34	Wang Y, Chen X, Cao W and Shi Y: Plasticity of mesenchymal stem cells in immunomodulation: Pathological and therapeutic implications. Nat Immunol. 15:1009–1016. 2014. View Article : Google Scholar : PubMed/NCBI
35	Creemers EE, Tijsen AJ and Pinto YM: Circulating microRNAs: Novel biomarkers and extracellular communicators in cardiovascular disease? Circ Res. 110:483–495. 2012. View Article : Google Scholar : PubMed/NCBI
36	Wang H, Han L, Zhao G, Shen H, Wang P, Sun Z, Xu C, Su Y, Li G, Tong T and Chen J: hnRNP A1 antagonizes cellular senescence and senescence-associated secretory phenotype via regulation of SIRT1 mRNA stability. Aging cell. Sep 9–2016.(Epub ahead of print). View Article : Google Scholar
37	McCubbrey AL, Nelson JD, Stolberg VR, Blakely PK, McCloskey L, Janssen WJ, Freeman CM and Curtis JL: MicroRNA-34a negatively regulates efferocytosis by tissue macrophages in part via SIRT1. 196:1366–1375. 2016.
38	Brunet A, Sweeney LB, Sturgill JF, Chua KF, Greer PL, Lin Y, Tran H, Ross SE, Mostoslavsky R, Cohen HY, et al: Stress-dependent regulation of FOXO transcription factors by the SIRT1 deacetylase. Science. 303:2011–2015. 2004. View Article : Google Scholar : PubMed/NCBI
39	Motta MC, Divecha N, Lemieux M, Kamel C, Chen D, Gu W, Bultsma Y, McBurney M and Guarente L: Mammalian SIRT1 represses forkhead transcription factors. Cell. 116:551–563. 2004. View Article : Google Scholar : PubMed/NCBI
40	Zhao T, Li J and Chen AF: MicroRNA-34a induces endothelial progenitor cell senescence and impedes its angiogenesis via suppressing silent information regulator 1. Am J Physiol Endocrinol Metab. 299:E110–E116. 2010. View Article : Google Scholar : PubMed/NCBI
41	Xiong H, Pang J, Yang H, Dai M, Liu Y, Ou Y, Huang Q, Chen S, Zhang Z, Xu Y, et al: Activation of miR-34a/SIRT1/p53 signaling contributes to cochlear hair cell apoptosis: Implications for age-related hearing loss. Neurobiol Aging. 36:1692–1701. 2015. View Article : Google Scholar : PubMed/NCBI
42	Chan SR and Blackburn EH: Telomeres and telomerase. Philos Trans R Soc Lond B Biol Sci. 359:109–121. 2004. View Article : Google Scholar : PubMed/NCBI
43	de Lange T: Shelterin: The protein complex that shapes and safeguards human telomeres. Genes Dev. 19:2100–2110. 2005. View Article : Google Scholar : PubMed/NCBI
44	Mourkioti F, Kustan J, Kraft P, Day JW, Zhao MM, Kost-Alimova M, Protopopov A, DePinho RA, Bernstein D, Meeker AK and Blau HM: Role of telomere dysfunction in cardiac failure in Duchenne muscular dystrophy. Nat Cell Biol. 15:895–904. 2013. View Article : Google Scholar : PubMed/NCBI
45	Boe AE, Eren M, Murphy SB, Kamide CE, Ichimura A, Terry D, McAnally D, Smith LH, Miyata T and Vaughan DE: Plasminogen activator inhibitor-1 antagonist TM5441 attenuates Nomega-nitro-L-arginine methyl ester-induced hypertension and vascular senescence. Circulation. 128:2318–2324. 2013. View Article : Google Scholar : PubMed/NCBI
46	De Bonis ML, Ortega S and Blasco MA: SIRT1 is necessary for proficient telomere elongation and genomic stability of induced pluripotent stem cells. Stem Cell Reports. 2:690–706. 2014. View Article : Google Scholar : PubMed/NCBI