Akt3 is a target of miR-29c-3p and serves an important function in the pathogenesis of congenital heart disease

Chen,Tao; Li,Shu‑Jun; Chen,Bin; Huang,Qiong; Kong,Xiang‑Ying; Shen,Chen; Gu,Hai‑Tao; Wang,Xiao‑Wei

doi:10.3892/ijmm.2018.4008

Akt3 is a target of miR-29c-3p and serves an important function in the pathogenesis of congenital heart disease

Authors:
- Tao Chen
- Shu‑Jun Li
- Bin Chen
- Qiong Huang
- Xiang‑Ying Kong
- Chen Shen
- Hai‑Tao Gu
- Xiao‑Wei Wang
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Affiliations: Department of Cardiothoracic Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China, Department of Pediatrics, Children's Hospital of Anhui Medical University, Hefei, Anhui 230000, P.R. China
Published online on: November 30, 2018 https://doi.org/10.3892/ijmm.2018.4008
Pages: 980-992
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Our previous studies identified that the expression of microRNA‑29c (miR‑29c‑3p) was significantly increased in the serum of pregnant women carrying fetuses with congenital heart disease (CHD) compared with in that of normal pregnant women. However, the mechanism by which miR‑29c‑3p affects development of the embryonic heart remained unclear. The aim of the present study was to investigate the effect and potential molecular mechanism of miR‑29c‑3p overexpression on P19 cell proliferation, apoptosis and differentiation. miR‑29c‑3p‑overexpression and protein kinase Bγ (Akt3)‑knockdown cell lines were constructed using transfection technology. The function of miR‑29c‑3p and Akt3 in cardiomyocyte development was investigated by determining the proliferation, apoptosis and differentiation of P19 cells, which can differentiate into cardiomyocytes induced by dimethylsulfoxide. Bioinformatic analysis and luciferase assays were performed to explore the association between Akt3 and miR‑29c‑3p. The results of the present study revealed that miR‑29c‑3p overexpression and Akt3 knockdown suppressed proliferation, and promoted apoptosis and differentiation in P19 cells. Akt3 was also demonstrated to be a target of miR‑29c‑3p. Therefore, overexpression of miR‑29c‑3p may inhibit proliferation, and promote apoptosis and differentiation in P19 cells by inhibiting the expression of Akt3. miR‑29c‑3p may be a potential therapeutic target for the treatment of CHD.

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1	Wang J and Liu B: Exercise and congenital heart disease. Adv Exp Med Biol. 1000:95–101. 2017. View Article : Google Scholar : PubMed/NCBI
2	Junghare SW and Desurkar V: Congenital heart diseases and anaesthesia. Indian J Anaesth. 61:744–752. 2017. View Article : Google Scholar : PubMed/NCBI
3	Itani M, Matesan M, Ahuja J, Bermo M, Habib AS, Goiney C, Krieger EV and Vesselle H: The role of pulmonary scintigraphy in the evaluation of adults with congenital heart disease. Semin Nucl Med. 47:660–670. 2017. View Article : Google Scholar
4	Suradi HS and Hijazi ZM: Adult congenital interventions in heart failure. Interv Cardiol Clin. 6:427–443. 2017.PubMed/NCBI
5	Chien PS, Chiang CB, Wang Z and Chiou TJ: MicroRNA-mediated signaling and regulation of nutrient transport and utilization. Curr Opin Plant Biol. 39:73–79. 2017. View Article : Google Scholar : PubMed/NCBI
6	de Lucia C, Komici K, Borghetti G, Femminella GD, Bencivenga L, Cannavo A, Corbi G, Ferrara N, Houser SR, Koch WJ and Rengo G: microRNA in cardiovascular aging and age-related cardiovascular diseases. Front Med (Lausanne). 4:742017. View Article : Google Scholar
7	Stanley-Hasnain S, Hauck L, Grothe D and Billia F: Control of cardiomyocyte proliferation through p53/Mdm2-regulated MicroRNAs. J Heart Lung Transpl. 35(Suppl 1): S183–S184. 2016. View Article : Google Scholar
8	Zhou Y, Jia WK, Jian Z, Zhao L, Liu CC, Wang Y and Xiao YB: Downregulation of microRNA-199a-5p protects cardiomyocytes in cyanotic congenital heart disease by attenuating endoplasmic reticulum stress. Mol Med Rep. 16:2992–3000. 2017. View Article : Google Scholar : PubMed/NCBI
9	Zhu S, Cao L, Zhu J, Kong L, Jin J, Qian L, Zhu C, Hu X, Li M, Guo X, et al: Identification of maternal serum MicroRNAs as novel noninvasive biomarkers for prenatal detection of fetal congenital heart defects. Clin Chim Acta. 424:66–72. 2013. View Article : Google Scholar : PubMed/NCBI
10	Shu YJ, Bao RF, Jiang L, Wang Z, Wang XA, Zhang F, Liang HB, Li HF, Ye YY, Xiang SS, et al: MicroRNA-29c-5p suppresses gallbladder carcinoma progression by directly targeting CPEB4 and inhibiting the MAPK pathway. Cell Death Differ. 24:445–457. 2017. View Article : Google Scholar :
11	Han TS, Hur K, Xu G, Choi B, Okugawa Y, Toiyama Y, Oshima H, Oshima M, Lee HJ, Kim VN, et al: MicroRNA-29c mediates initiation of gastric carcinogenesis by directly targeting ITGB1. Gut. 64:203–214. 2015. View Article : Google Scholar
12	Zhang HW, Wang EW, Li LX, Yi SH, Li LC, Xu FL, Wang DL, Wu YZ and Nian WQ: A regulatory loop involving miR-29c and Sp1 elevates the TGF-β1 mediated epithelial-to-mesenchymal transition in lung cancer. Oncotarget. 7:85905–85916. 2016.PubMed/NCBI
13	Niu M, Gao D, Wen Q, Wei P, Pan S, Shuai C, Ma H, Xiang J, Li Z, Fan S, et al: MiR-29c regulates the expression of miR-34c and miR-449a by targeting DNA methyltransferase 3a and 3b in nasopharyngeal carcinoma. BMC Cancer. 16:2182016. View Article : Google Scholar : PubMed/NCBI
14	Butrym A, Rybka J, Baczyńska D, Poręba R, Kuliczkowski K and Mazur G: Clinical response to azacitidine therapy depends on microRNA-29c (miR-29c) expression in older acute myeloid leukemia (AML) patients. Oncotarget. 7:30250–30257. 2016. View Article : Google Scholar : PubMed/NCBI
15	Nygren MK, Tekle C, Ingebrigtsen VA, Mäkelä R, Krohn M, Aure MR, Nunes-Xavier CE, Perälä M, Tramm T, Alsner J, et al: Identifying microRNAs regulating B7-H3 in breast cancer: The clinical impact of microRNA-29c. Br J Cancer. 110:2072–2080. 2014. View Article : Google Scholar : PubMed/NCBI
16	Wang S, Zhu Y and Qiu R: Shikonin protects H9C2 cardiomyocytes against hypoxia/reoxygenation injury through activation of PI3K/Akt signaling pathway. Biomed Pharmacother. 104:712–717. 2018. View Article : Google Scholar
17	Taniyama Y, Ito M, Sato K, Kuester C, Veit K, Tremp G, Liao R, Colucci WS, Ivashchenko Y, Walsh K and Shiojima I: Akt3 over-expression in the heart results in progression from adaptive to maladaptive hypertrophy. J Mol Cell Cardiol. 38:375–385. 2005. View Article : Google Scholar : PubMed/NCBI
18	Shiojima I, Sato K, Izumiya Y, Schiekofer S, Ito M, Liao R, Colucci WS and Walsh K: Disruption of coordinated cardiac hypertrophy and angiogenesis contributes to the transition to heart failure. J Clin Invest. 115:2108–2118. 2005. View Article : Google Scholar :
19	Yoshioka K, Yoshida K, Cui H, Wakayama T, Takuwa N, Okamoto Y, Du W, Qi X, Asanuma K, Sugihara K, et al: Endothelial PI3K-C2α, a class II PI3K, has an essential role in angiogenesis and vascular barrier function. Net Med. 18:1560–1569. 2012. View Article : Google Scholar
20	Kanungo J: Retinoic acid signaling in P19 stem cell differentiation. Anticancer Agents Med Chem. 17:1184–1198. 2017. View Article : Google Scholar
21	Wang L, Song G, Liu M, Chen B, Chen Y, Shen Y, Zhu J and Zhou X: MicroRNA-375 overexpression influences P19 cell proliferation, apoptosis and differentiation through the notch signaling pathway. Int J Mol Med. 37:47–55. 2016. View Article : Google Scholar
22	McBurney MW, Jones-Villeneuve EM, Edwards MK and Anderson PJ: Control of muscle and neuronal differentiation in a cultured embryonal carcinoma cell line. Nature. 299:165–167. 1982. View Article : Google Scholar
23	Mueller I, Kobayashi R, Nakajima T, Ishii M and Ogawa K: Effective and steady differentiation of a clonal derivative of P19CL6 embryonal carcinoma cell line into beating cardiomyocytes. J Biomed Biotechnol. 2010:3805612010. View Article : Google Scholar
24	Song G, Shen Y, Ruan Z, Li X, Chen Y, Yuan W, Ding X, Zhu L and Qian L: LncRNA-uc 167 influences cell proliferation, apoptosis and differentiation of P19 cells by regulating Mef2c. Gene. 590:97–108. 2016. View Article : Google Scholar : PubMed/NCBI
25	van der Heyden MA and Defize LH: Twenty one years of P19 cells: What an embryonal carcinoma cell line taught us about cardiomyocyte differentiation. Cardiovasc Res. 58:292–302. 2003. View Article : Google Scholar
26	Mcburney MW: P19 embryonal carcinoma cells. Int J Dev Biol. 37:135–140. 1993.
27	Ai F, Zhang Y and Peng B: miR-20a regulates proliferation, differentiation and apoptosis in P19 cell model of cardiac differentiation by targeting smoothened. Biol Open. 5:1260–1265. 2016. View Article : Google Scholar
28	Wang D, Liu C, Wang Y, Wang W, Wang K, Wu X, Li Z, Zhao C, Li L and Peng L: Impact of miR-26b on cardiomyocyte differentiation in P19 cells through regulating canonical/non-canonical wnt signaling. Cell Proliferation. 50:e123712017. View Article : Google Scholar
29	Li H, Jiang L, Yu Z, Han S, Liu X, Li M, Zhu C, Qiao L and Huang L: The role of a novel long noncoding RNA TUC40- in cardiomyocyte induction and maturation in P19 cells. Am J Med Sci. 354:608–616. 2017. View Article : Google Scholar : PubMed/NCBI
30	Van Poucke M, Van Zeveren A and Peelman LJ: [Letter to the editor] Combined FAM-labeled TaqMan probe detection and SYBR green I melting curve analysis in multiprobe qPCR genotyping assays. Biotechniques. 52:81–86. 2012. View Article : Google Scholar
31	Wang J, Meng X, Dobrovolskaya OB, Orlov YL and Chen M: Non-coding RNAs and their roles in stress response in plants. Genomics Proteomics Bioinformatics. 15:301–312. 2017. View Article : Google Scholar : PubMed/NCBI
32	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
33	Grossi I, Salvi A, Abeni E, Marchina E and De Petro G: Biological function of MicroRNA193a-3p in health and disease. Int J Genomics. 2017:59131952017. View Article : Google Scholar : PubMed/NCBI
34	Kumar V, Khare T, Shriram V and Wani SH: Plant small RNAs: The essential epigenetic regulators of gene expression for salt-stress responses and tolerance. Plant Cell Rep. 37:61–75. 2018. View Article : Google Scholar
35	Chen C, Ponnusamy M, Liu C, Gao J, Wang K and Li P: MicroRNA as a therapeutic target in cardiac remodeling. Biomed Res Int. 2017:12784362017. View Article : Google Scholar : PubMed/NCBI
36	Wojciechowska A, Braniewska A and Kozar-Kamińska K: MicroRNA in cardiovascular biology and disease. Adv Clin Exp Med. 26:865–874. 2017. View Article : Google Scholar
37	Sun W, Zhao L, Song X, Zhang J, Xing Y, Liu N, Yan Y, Li Z, Lu Y, Wu J, et al: MicroRNA-210 modulates the cellular energy metabolism shift during H2O2-induced oxidative stress by repressing ISCU in H9c2 cardiomyocytes. Cell Physiol Biochem. 43:383–394. 2017. View Article : Google Scholar
38	Chu D, Zhao Z, Li Y, Li J, Zheng J, Wang W, Zhao Q and Ji G: Increased microRNA-630 expression in gastric cancer is associated with poor overall survival. PLoS One. 9:e905262014. View Article : Google Scholar : PubMed/NCBI
39	Qiao P, Li G, Bi W, Yang L, Yao L and Wu D: microRNA-34a inhibits epithelial mesenchymal transition in human cholangio-carcinoma by targeting Smad4 through transforming growth factor-beta/Smad pathway. BMC Cancer. 15:4692015. View Article : Google Scholar
40	Rijlaarsdam MA and Looijenga LH: An oncofetal and developmental perspective on testicular germ cell cancer. Semin Cancer Biol. 29:59–74. 2014. View Article : Google Scholar : PubMed/NCBI
41	Paraskevas G, Koutsouflianiotis K and Iliou K: The first descriptions of various anatomical structures and embryological remnants of the heart: A systematic overview. Int J Cardiol. 227:674–690. 2017. View Article : Google Scholar
42	Gittenbergerde Groot AC, Bartelings MM, Poelmann RE, Haak MC and Jongbloed MR: Embryology of the heart and its impact on understanding fetal and neonatal heart disease. Semin Fetal Neonatal Med. 18:237–244. 2013. View Article : Google Scholar
43	Abu-Elmagd M and Wheeler G: Frizzled-7 expression during early cardiogenesis of Xenopus laevis embryo. BMC Genomics. 15:1–2. 2014. View Article : Google Scholar
44	Vitagliano O, Addeo R, D’Angelo V, Indolfi C, Indolfi P and Casale F: The Bcl-2/Bax and Ras/Raf/MEK/ERK signaling pathways: Implications in pediatric leukemia pathogenesis and new prospects for therapeutic approaches. Expert Rev Hematol. 6:587–597. 2013. View Article : Google Scholar : PubMed/NCBI
45	Siddiqui WA, Ahad A and Ahsan H: The mystery of BCL2 family: Bcl-2 proteins and apoptosis: An update. Arch Toxicol. 89:289–317. 2015. View Article : Google Scholar : PubMed/NCBI
46	Reed JC: Proapoptotic multidomain Bcl-2/Bax-family proteins: Mechanisms, physiological roles, and therapeutic opportunities. Cell Death Differ. 13:1378–1386. 2006. View Article : Google Scholar : PubMed/NCBI