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| Effect of miRNA-1 and miRNA-133 overexpression on myocardial differentiation of inducible pluripotent stem cells |
| WANG Fangfang BAI Baobao▲ ZENG Di CHU Yi LI Xue |
| Department of Cardiology, the Second Affiliated Hospital of PLA Air Force Military Medical University, Shaanxi Province, Xi′an 710038, China |
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Abstract Objective To investigate the effect of miRNA-1 and miRNA-133 overexpression on myocardial differentiation of inducible pluripotent stem cells (iPSC). Methods Embryos from two-week-old American Institute of Cancer Research (ICR) pregnant mice were collected, 0.5 g/L of trypsin was used for digestion of vaccination, mitomycin was used after spreading to three generations, mouse induced pluripotent stem cells (miPSC) was incubated and cultivated at 37℃, miPSC growed on embryonic fibroblasts was digested and passaged. Medium was replaced, miRNA (U6) (10 nmol/L, marked as control group), miRNA-1 (10 nmol/L, marked as miRNA-1 group), miRNA-133 (10 nmol/L, marked as miRNA-133 group), miRNA-1 and miRNA-133 (10 nmol/L respectively, marked as miRNA-1+133 group) were added and transfected for 24 hours, and were washed by PBS, then miPSC differentiation medium was added, and its differentiation was daily observed, cell pulse rate was calculated. RT-PCR was used to detect the expression of miRNA-1, miRNA-133 and cardiac troponin T (cTnT). Results The cTnT was barely expressed in undifferentiated miPSC, compared with 0 day of differentiation, the expreesion level of cTnT of miRNA-1 group and miRNA-133 group was increased after 5 days of differentiation (P ＜ 0.05), the increase was more obvious after 10 days of differentiation (P ＜ 0.01). After 10 days of differentiation, the expression level of cTnT of miRNA-1+133 group was obviously higher than those of miRNA-1 group and miRNA-133 group (P ＜ 0.01). Compared with control group, the cell pulse rate of miRNA-1 group and miRNA-133 group were not increased (P ＞ 0.05), while the cell pulse rate of miRNA-1+133 group was significantly increased (P ＜ 0.01). Conclusion miRNA-1 or miRNA-133 alone does not promote myocardial differentiation, only after the co-expression of miRNA-1 and miRNA-133, it can significantly improve the directional differentiation efficiency of miPSC to cardiac cells.
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[1] Tu Y，Sun Y，Fan Y，et al. Multimodality Molecular Imaging of Cardiovascular Disease Based on Nanoprobes [J]. Cell Physiol Biochem，2018，48（4）：1401-1415.
[2] Coban M，Inci A，Y?覦lmaz U，et al. The Association of Fibroblast Growth Factor 23 with Arterial Stiffness and Ath-erosclerosis in Patients with Autosomal Dominant Polycystic Kidney Disease [J]. Kidney Blood Press Res，2018，43（4）：1160-1173.
[3] Matteucci C，Argaw-Denboba A，Balestrieri E，et al. Deciphering cellular biological processes to clinical application：a new perspective for Tα1 treatment targeting multiple diseases [J]. Expert Opin Biol Ther，2018，18（sup1）：23-31.
[4] Mahapatra S，Martin D，Gallicano GI. Re-Defining Stem Cell-Cardiomyocyte Interactions：Focusing on the Paracrine Effector Approach [J]. J Stem Cells Regen Med，2018，14（1）：10-26.
[5] Xie A，Zhou A，Liu H，et al. Mitochondrial Ca2+ flux modulates spontaneous electrical activity in ventricular cardiomyocytes [J]. PLoS One，2018，13（7）：e0200448.
[6] Abou-Saleh H，Zouein FA，El-Yazbi A，et al. The march of pluripotent stem cells in cardiovascular regenerative medicine [J]. Stem Cell Res Ther，2018，9（1）：201.
[7] Garg P，Garg V，Shrestha R，et al. Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes as Models for Cardiac Channelopathies：A Primer for Non-Electrophysiologists [J]. Circ Res，2018，123（2）：224-243.
[8] Ziegler T，Hinkel R，Kupatt C. Induced pluripotent stem cell derived cardiac models：effects of Thymosin β4 [J]. Expert Opin Biol Ther，2018，18（sup1）：111-120.
[9] Prajapati C，P?觟l?觟nen RP，Aalto-Set?覿l?覿 K. Simultaneous recordings of action potentials and calcium transients from human induced pluripotent stem cell derived cardiomyocytes [J]. Biol Open，2018，7（7）：bio035030.
[10] Duan F，Huang R，Zhang F，et al. Biphasic modulation of insulin signaling enables highly efficient hematopoietic differentiation from human pluripotent stem cells [J]. Stem Cell Res Ther，2018，9（1）：205.
[11] Xu H，He JH，Xu SJ，et al. A group of tissue-specific microRNAs contribute to the silencing of CUX1 in different cell lineages during development [J]. J Cell Biochem，2018，119（7）：6238-6248.
[12] Alfar EA，El-Armouche A，Guan K. MicroRNAs in cardiomyocyte differentiation and maturation [J]. Cardiovasc Res，2018，114（6）：779-781.
[13] 祝烨，宋鑫.miRNA与lncRNA的相互调控作用在肿瘤中的研究进展[J].基础医学与临床，2015，35（11）：1554-1558.
[14] Poon EN，Hao B，Guan D，et al. Integrated transcriptomic and regulatory network analyses identify microRNA-200c as a novel repressor of human pluripotent stem cell-derived cardiomyocyte differentiation and maturation [J]. Cardiovasc Res，2018，114（6）：894-906.
[15] Ling X，Yao D，Kang L，et al. Involment of RAS/ERK1/2 signaling and MEF2C in miR-155-3p inhibition-triggered cardiomyocyte differentiation of embryonic stem cell [J]. Oncotarget，2017，8（48）：84403-84416.
[16] 黄辰，刘利英，倪磊，等.肿瘤miRNAs调控机制的研究进展与展望[J].西安交通大学学报：医学版，2015，36（4）：429-434.
[17] 汤志雄，苟德明.miRNA调控成肌分化的研究进展[J].中国生物工程杂志，2017，37（10）：103-110.
[18] Guo X，Bai Y，Zhang L，et al. Cardiomyocyte differentiation of mesenchymal stem cells from bone marrow：new regulators and its implications [J]. Stem Cell Res Ther，2018，9（1）：44.
[19] 戴备军，张美齐，翁剑枫，等.miRNA-1和心肺复苏后心功能障碍严重程度的相关性研究[J].中国现代医生，2017，55（11）：5-7，10.
[20] Nikolova-Krstevski V，Wagner S，Yu ZY，et al. Endocardial TRPC-6 Channels Act as Atrial Mechanosensors and Load-Dependent Modulators of Endocardial/Myocardial Cross-Talk [J]. JACC Basic Transl Sci，2017，2（5）：575-590.
[21] Ma D，Liu Z，Loh LJ，et al. Identification of an INa-dependent and Ito-mediated proarrhythmic mechanism in cardiomyocytes derived from pluripotent stem cells of a Brugada syndrome patient [J]. Sci Rep，2018，8（1）：11246.
[22] Takaya T，Ono K，Kawamura T，et al. MicroRNA-1 and MicroRNA-133 in spontaneous myocardial differentiation of mouse embryonic stem cells [J]. Circ J，2009，73（8）：1492-1497. |
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