Rapamycin‑induced miR‑30a downregulation inhibits senescence of VSMCs by targeting Beclin1

Tan,Pan; Wang,Haiqin; Zhan,Junkun; Ma,Xinyu; Cui,Xingjun; Wang,Yanjiao; Wang,Yi; Zhong,Jiayu; Liu,Youshuo

doi:10.3892/ijmm.2019.4074

Rapamycin‑induced miR‑30a downregulation inhibits senescence of VSMCs by targeting Beclin1

Authors:
- Pan Tan
- Haiqin Wang
- Junkun Zhan
- Xinyu Ma
- Xingjun Cui
- Yanjiao Wang
- Yi Wang
- Jiayu Zhong
- Youshuo Liu
View Affiliations

Affiliations: Department of Geriatrics, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China, Department of Anesthesiology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China
Published online on: January 23, 2019 https://doi.org/10.3892/ijmm.2019.4074
Pages: 1311-1320
Copyright: © Tan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Vascular senescence is considered to be an independent risk factor for cardiovascular diseases. The present study aimed to investigate the effects of rapamycin on miR‑30a and its relationship with autophagy and senescence in vascular smooth muscle cells (VSMCs). Young and aging VSMCs were treated with rapamycin or transfected with miR‑30a mimics. Measurement of cellular senescence was conducted using senescence‑associated (SA)‑β‑Galactosidase (gal) staining. Dual luciferase reporter assay was used to confirm binding for miR‑30a and Beclin1. The expression levels of miR‑30a and Beclin1 were determined with reverse transcription‑quantitative polymerase chain reaction analysis. Autophagy‑related protein levels were determined using immunofluorescence or western blot assays. The results demonstrated that rapamycin treatment significantly decreased miR‑30a expression and increased Beclin1 expression in both young and aging cells, as well as promoted autophagy in VSMCs. In addition, rapamycin inhibited senescence in VSMCs and could also alleviate the aging VSMC cycle arrest. Dual luciferase reporter assay confirmed that miR‑30a could directly bind the 3'untranslated region of Beclin1 and inhibit its expression. Furthermore, miR‑30a inhibited autophagy and promoted senescence of VSMCs. In conclusion, the present results indicated that rapamycin could inhibit the senescence of VSMCs by downregulating miR‑30a, which resulted in upregulation of Beclin1 and activation of autophagy. The current study is the first to demonstrate an inhibitory role of rapamycin on VSMC senescence and might provide novel insights and potential new molecular targets in senescence treatment.

View Figures

View References

1	Sueta D, Koibuchi N, Yu H, Toyama K, Uekawa K, Katayama T, Ma MJ, Nakagawa T, Waki H, Maeda M, et al: Blood pressure variability, impaired autonomic function and vascular senescence in aged spontaneously hypertensive rats are ameliorated by angiotensin blockade. Atherosclerosis. 236:101–107. 2014. View Article : Google Scholar : PubMed/NCBI
2	Rodier F and Campisi J: Four faces of cellular senescence. J Cell Biol. 192:547–556. 2011. View Article : Google Scholar : PubMed/NCBI
3	Wang J, Uryga AK, Reinhold J, Figg N, Baker L, Finigan A, Gray K, Kumar S, Clarke M and Bennett M: Vascular smooth muscle cell senescence promotes atherosclerosis and features of plaque vulnerability. Circulation. 132:1909–1919. 2015. View Article : Google Scholar : PubMed/NCBI
4	Mistry Y, Poolman T, Williams B and Herbert KE: A role for mitochondrial oxidants in stress-induced premature senescence of human vascular smooth muscle cells. Redox Biol. 1:411–417. 2013. View Article : Google Scholar : PubMed/NCBI
5	Campisi J: Aging, cellular senescence, and cancer. Annu Rev Physiol. 75:685–705. 2013. View Article : Google Scholar
6	Birch J, Barnes PJ and Passos JF: Mitochondria, telomeres and cell senescence: Implications for lung ageing and disease. Pharmacol Ther. 183:34–49. 2018. View Article : Google Scholar
7	Gewirtz DA: Autophagy and senescence: A partnership in search of definition. Autophagy. 9:808–812. 2013. View Article : Google Scholar : PubMed/NCBI
8	Gewirtz DA: Autophagy and senescence in cancer therapy. J Cell Physiol. 229:6–9. 2014.
9	Tan P, Wang YJ, Li S, Wang Y, He JY, Chen YY, Deng HQ, Huang W, Zhan JK and Liu YS: The PI3K/Akt/mTOR pathway regulates the replicative senescence of human VSMCs. Mol Cell Biochem. 422:1–10. 2016. View Article : Google Scholar : PubMed/NCBI
10	Wong M, Leung L and Nighot PK: Role of autophagy related protein ATG6/Beclin 1 in intestinal tight junction barrier. Gastroenterology. 152(Suppl 1): S1192017. View Article : Google Scholar
11	Kim J and Guan KL: Regulation of the autophagy initiating kinase ULK1 by nutrients: Roles of mTORC1 and AMPK. Cell Cycle. 10:1337–1338. 2011. View Article : Google Scholar : PubMed/NCBI
12	Pattingre S, Espert L, Biardpiechaczyk M and Codogno P: Regulation of macroautophagy by mTOR and Beclin1 complexes. Biochimie. 90:313–323. 2008. View Article : Google Scholar
13	Zhan JK, Wang YJ, Wang Y, Wang S, Tan P, Huang W and Liu YS: The mammalian target of rapamycin signalling pathway is involved in osteoblastic differentiation of vascular smooth muscle cells. Can J Cardiol. 30:568–575. 2014. View Article : Google Scholar : PubMed/NCBI
14	Wilkinson JE, Burmeister L, Brooks SV, Chan CC, Friedline S, Harrison DE, Hejtmancik JF, Nadon N, Strong R, Wood LK, et al: Rapamycin slows aging in mice. Aging Cell. 11:675–682. 2012. View Article : Google Scholar : PubMed/NCBI
15	Pospelova TV, Leontieva OV, Bykova TV, Zubova SG, Pospelov VA and Blagosklonny MV: Suppression of replicative senescence by rapamycin in rodent embryonic cells. Cell Cycle. 11:2402–2407. 2012. View Article : Google Scholar : PubMed/NCBI
16	Rong W, Zhen Y, Sunchu B, Caples K, Zhao S, Dang I and Perez VI: Rapamycin modulates cell senescence and inflammation by different mechanisms. Exp Gerontol. 94:126–127. 2017. View Article : Google Scholar
17	Muther C, Jobeili L, Garion M, Heraud S, Thepot A, Damour O and Lamartine J: An expression screen for aged-dependent microRNAs identifies miR-30a as a key regulator of aging features in human epidermis. Aging (Albany NY). 9:2376–2396. 2017. View Article : Google Scholar :
18	Lin X, Zhan JK, Wang YJ, Tan P, Chen YY, Deng HQ and Liu YS: Function, role, and clinical application of MicroRNAs in vascular aging. Biomed Res Int. 2016:60213942016. View Article : Google Scholar
19	Zhu H, Wu H, Liu X, Li B, Chen Y, Ren X, Liu CG and Yang JM: Regulation of autophagy by a beclin 1-targeted microRNA, miR-30a, in cancer cells. Autophagy. 5:816–823. 2009. View Article : Google Scholar : PubMed/NCBI
20	Xu R, Liu S, Chen H and Lao L: MicroRNA-30a downregulation contributes to chemoresistance of osteosarcoma cells through activating Beclin-1-mediated autophagy. Oncology Reports. 35:1757–1763. 2016. View Article : Google Scholar
21	Lee KY, Kim JR and Choi HC: Genistein-induced LKB1-AMPK activation inhibits senescence of VSMC through autophagy induction. Vascul Pharmacol. 81:75–82. 2016. View Article : Google Scholar : PubMed/NCBI
22	Zhang Y, Qian X, Sun X, Lin C, Jing Y, Yao Y, Ma Z, Kuai M, Lu Y, Kong X, et al: Liuwei Dihuang, a traditional Chinese medicinal formula, inhibits proliferation and migration of vascular smooth muscle cells via modulation of estrogen receptors. Int J Mol Med. 42:31–40. 2018.PubMed/NCBI
23	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
24	Luo Y, Li L, Zou P, Wang J, Shao L, Zhou D and Liu L: Rapamycin enhances long-term hematopoietic reconstitution of ex vivo expanded mouse hematopoietic stem cells by inhibiting senescence. Transplantation. 97:20–29. 2014. View Article : Google Scholar
25	Hinojosa CA, Mgbemena V, Van Roekel S, Austad SN, Miller RA, Bose S and Orihuela CJ: Enteric-delivered rapamycin enhances resistance of aged mice to pneumococcal pneumonia through reduced cellular senescence. Exp Gerontol. 47:958–965. 2012. View Article : Google Scholar : PubMed/NCBI
26	Blagosklonny MV: Progeria, rapamycin and normal aging: Recent breakthrough. Aging (Albany NY). 3:685–691. 2011. View Article : Google Scholar
27	Qi LW, Zhang Z, Zhang CF, Anderson S, Liu Q, Yuan CS and Wang CZ: Anti-colon cancer effects of 6-Shogaol through G2/M cell cycle arrest by p53/p21-cdc2/cdc25A crosstalk. Am J Chin Med. 43:743–756. 2015. View Article : Google Scholar : PubMed/NCBI
28	Wan D, Jiang C, Hua X, Wang T and Chai Y: Cell cycle arrest and apoptosis induced by aspidin PB through the p53/p21 and mitochondria-dependent pathways in human osteosarcoma cells. Anticancer Drugs. 26:931–941. 2015. View Article : Google Scholar : PubMed/NCBI
29	Sotthibundhu A, McDonagh K, von Kriegsheim A, Garcia-Munoz A, Klawiter A, Thompson K, Chauhan KD, Krawczyk J, Mcinerney V, Dockery P, et al: Rapamycin regulates autophagy and cell adhesion in induced pluripotent stem cells. Stem Cell Res Ther. 7:1662016. View Article : Google Scholar : PubMed/NCBI
30	Russell RC, Tian Y, Yuan H, Park HW, Chang YY, Kim J, Kim H, Neufeld TP, Dillin A and Guan KL: ULK1 induces autophagy by phosphorylating Beclin-1 and activating VPS34 lipid kinase. Nat Cell Biol. 15:741–750. 2013. View Article : Google Scholar : PubMed/NCBI
31	Xu J, Fotouhi M and McPherson PS: Phosphorylation of the exchange factor DENND3 by ULK in response to starvation activates Rab12 and induces autophagy. EMBO Rep. 16:709–718. 2015. View Article : Google Scholar : PubMed/NCBI
32	Chiao YA, Kolwicz SC, Basisty N, Gagnidze A, Zhang J, Gu H, Djukovic D, Beyer RP, Raftery D, MacCoss MJ, et al: Rapamycin transiently induces mitochondrial remodeling to reprogram energy metabolism in old hearts. Aging (Albany NY). 8:314–327. 2016. View Article : Google Scholar
33	Sarbassov DD, Ali SM, Kim DH, Guertin DA, Latek RR, Erdjument-Bromage H, Tempst P and Sabatini DM: Rictor, a novel binding partner of mTOR, defines a rapamycin-insensitive and raptor-independent pathway that regulates the cytoskeleton. Curr Biol. 14:1296–1302. 2004. View Article : Google Scholar : PubMed/NCBI
34	Huang M, Ke Y, Sun X, Yu L, Yang Z, Zhang Y, Du M, Wang J, Liu X and Huang SJ: Mammalian target of rapamycin signaling is involved in the vasculogenic mimicry of glioma via hypoxia-inducible factor-1α. Oncol Rep. 32:1973–1980. 2014. View Article : Google Scholar : PubMed/NCBI
35	Gu Z, Tan W, Ji J, Feng G, Meng Y, Da Z, Guo G, Xia Y, Zhu X, Shi G and Cheng C: Rapamycin reverses the senescent phenotype and improves immunoregulation of mesenchymal stem cells from MRL/lpr mice and systemic lupus erythematosus patients through inhibition of the mTOR signaling pathway. Aging (Albany NY). 8:1102–1114. 2016. View Article : Google Scholar
36	Fu XT, Shi YH, Zhou J, Peng YF, Liu WR, Shi GM, Gao Q, Wang XY, Song K, Fan J and Ding ZB: MicroRNA-30a suppresses autophagy-mediated anoikis resistance and metastasis in hepatocellular carcinoma. Cancer Lett. 412:108–117. 2018. View Article : Google Scholar
37	Zhang L, Cheng R and Huang Y: MiR-30a inhibits BECN1-mediated autophagy in diabetic cataract. Oncotarget. 8:77360–77368. 2017.PubMed/NCBI
38	Geng GJ, Yang YT, Jiang J, Yu XY and Fa XE: MicroRNA-30a suppresses non-small-cell lung cancer by targeting Myb-related protein B. Exp Ther Med. 15:1633–1639. 2018.PubMed/NCBI
39	Wang X, Xu X, Wei W, Yu Z, Wen L, He K and Fan H: MicroRNA-30a-5p promotes replication of porcine circovirus type 2 through enhancing autophagy by targeting 14-3-3. Arch Virol. 162:2643–2654. 2017. View Article : Google Scholar : PubMed/NCBI
40	Zou Z, Wu L, Ding H, Wang Y, Zhang Y, Chen X, Chen X, Zhang CY, Zhang Q and Zen K: MicroRNA-30a sensitizes tumor cells to cis-platinum via suppressing beclin 1-mediated Autophagy. J Biol Chem. 287:4148–4156. 2012. View Article : Google Scholar :
41	Wang P, Liang J, Li Y and Li J, Yang X, Zhang X, Han S, Li S and Li J: Down-regulation of miRNA-30a alleviates cerebral isch-emic injury through enhancing beclin 1-mediated autophagy. Neurochem Res. 39:1279–1291. 2014. View Article : Google Scholar : PubMed/NCBI