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Journal of Peking University(Health Sciences) ›› 2019, Vol. 51 ›› Issue (2): 221-227. doi: 10.19723/j.issn.1671-167X.2019.02.004

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Role of miR-106b-5p in the regulation of gene profiles in endothelial cells

Jing ZHANG,Su-fang LI,Hong CHEN(),Jun-xian SONG   

  1. Center for Cardiovascular Translational Research, Peking University People’s Hospital, Beijing 100044, China;
  • Received:2017-05-22 Online:2019-04-18 Published:2019-04-26
  • Contact: Hong CHEN E-mail:chenhongbj@medmail.com.cn
  • Supported by:
    the National Natural Science Foundation of China(81400264)

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Abstract:

Objective: To evaluate the role of miR-106b-5p in the regulation of gene expression in endothelial cells.Methods: The Taqman low-density microRNAs (miRNAs) array (TLDA) was used to identify miRNA expression profiles in the plasma of patients with atherosclerotic coronary artery disease (CAD) (atherosclerosis group, n=9) and individuals without atherosclerotic CAD disease (control group, n=9). A weighed and undirected miRNA coexpression network analysis was performed to investigate the interactions among miRNAs in the two groups. MiR-106b-5p, whose coexpression pattern in atherosclerosis group was most different from that of control group, was further studied. Human umbilical vein endothelial cells (HUVEC) were transfected with miR-106b-5p mimic or negative control mimic, and Affymetrix GeneChip Human Transcriptome Array 2.0 was used to screen the differential gene expression profiles after transfection. And the signal transduction pathway of differential gene profiles was further analyzed in Kyoto Encyclopedia of Genes and Genomes (KEGG) signal pathway database. After parsing the whole KEGG database, all differentially expressed genes involved pathways were extracted, and the hypergeometric distribution was used to calculate the pathway enrichment.Results: The coexpression pattern of the patients with atherosclerosis (140 nodes, 1 154 edges) differed from that of the non-atherosclerosis control group (140 nodes, 612 edges). The analysis of array data with significant analysis of microarray (SAM) identified 746 significantly deregulated genes (fold change ≥ 1.5 and false discovery rate < 0.01) altered by overexpression of miR-106b-5p with miR-106b-5p mimic in HUVEC. By calculating the pathway enrichment, we found that multiple signaling pathways enriched in differential gene profiles were closely related to the process of formation and rupture of atherosclerotic plaque, including phosphatidylinositol-3 kinase (PI3K)/ protein kinase B (PKB, also called Akt), mammalian target of rapamycin (mTOR), transforming growth factor-β (TGF-β), janus kinase / signal transducer and activator of transcription (Jak-STAT), tumor necrosis factor (TNF), toll like receptor (TLR) and hypoxia-inducible factor 1α (HIF-1α) and other signal pathways.Conclusion: The coexpression pattern of miRNAs in plasma of patients with atherosclerosis is more significantly changed than that of individuals without atherosclerotic disease. MiR-106b-5p, which shows the most significant dif-ference between groups, targets multiple signal pathways in vascular endothelial cells, and might play an important role in the regulatory network of atherosclerotic gene expression.

Key words: Atherosclerosis, MicroRNAs, Transcriptome, Endothelial cells

CLC Number: 

  • R541.4

Table 1

Clinical characteristics of the study population for plasma miRNAs profiling"

Items Control group (n=9) Atherosclerosis group (n=9) P value
Gender, Male/Female 5/4 5/4 >0.99
Age/years, x?±s 56.1±9.3 60.4±9.0 0.33
BMI/(kg/m2), x?±s 24.5±2.3 24.8±4.0 0.86
PLT/(×1012/L), x?±s 219.2±79.3 209.3±46.2 0.75
WBC/(×109/L), x?±s 6.9±2.8 5.8±1.5 0.31
LDL cholesterol/(mmol/l), x?±s 2.5±0.9 2.4±0.5 0.65
Hypertension, n(%) 5 (55.6) 8 (88.9) 0.11
Diabetes mellitus, n(%) 1 (11.1) 2 (22.2) 0.53
Hyperlipemia, n(%) 5 (55.6) 6 (66.7) 0.63
CCB, n(%) 2 (22.2) 3 (33.3) 0.60
Beta-blocker, n(%) 3 (33.3) 5 (55.6) 0.34
Aspirin, n(%) 1 (11.1) 4 (44.4) 0.11
Clopidogrel, n(%) 0 3 (33.3) 0.06
ACEI, n(%) 1 (11.1) 2 (22.2) 0.53
ARB, n(%) 1 (11.1) 2 (22.2) 0.53

Figure 1

Coexpression network analysis of plasma miRNAs in controls (A) and atherosclerosis patients (B) Coexpression network constructed with all detected miRNAs. Nodes represent individual miRNAs, and edges represent coexpression relationships between miRNA pairs. The coexpression pattern of the atherosclerosis group (140 nodes, 1 154 edges) differed from that of the control group (141 nodes, 612 edges). miR-106b-5p was labeled with red node."

Table 2

miRNAs with differential coexpression pattern in the plasma of atherosclerosis patients compared to controls"

microRNA ID Degree in atherosclerosis Degree in controls K1-atherosclerosis K2-controls DiffK
miR-106-5p 43 7 1.00 0.21 0.78
miR-636 4 28 0.09 0.85 0.76
miR-302b 3 27 0.07 0.81 0.75
miR-15a* 3 24 0.07 0.73 0.66
miR-597 2 21 0.05 0.64 0.59
miR-543 31 5 0.72 0.15 0.57
miR-196b 15 29 0.35 0.88 0.53
miR-1271 6 22 0.14 0.67 0.53
miR-212 6 22 0.14 0.67 0.53
miR-29b 33 8 0.77 0.24 0.53
miR-365 25 2 0.58 0.06 0.52
miR-19b-1* 21 33 0.49 1.00 0.51
miR-452 16 29 0.37 0.88 0.51
miR-886-3p 27 4 0.63 0.12 0.51

Figure 2

Profile of mRNAs in human umbilical vein endothelial cells transfected with miR-106b-5p mimics compared with negative control mimic Heat map illustrates the levels of significantly changed mRNAs (fold change ≥ 1.5 and FDR<0.01) in human umbilical vein endothelial cells (HUVEC) transfected with miR-106b-5p mimics compared with negative control mimic. Color intensity is scaled within each row, such that the highest expression value corresponds to bright red and the lowest to bright green."

Figure 3

Potential gene signaling pathway targets of miR-106b-5p in human umbilical vein endothelial cells PI3K-Akt, phosphoinositide 3-kinase / protein kinase B; mTOR, mammalian target of rapamycin; Jak-STAT, janus kinase/signal transducer and activator of transcription; TGF-β, transforming growth factor-β; HIF-1, hypoxia-inducible factor 1; MAPK, mitogen-activated protein kinase; TNF, tumor necrosis factor; VEGF, vascular endothelial growth factor; ECM, extracellular matrix. Potential gene signaling pathway targets of miR-106b-5p in human umbilical vein endothelial cells (HUVEC) that showed significant difference between groups (HUVEC transfected with miR-106b-5p mimics compared with negative control mimic, P<0.05) were listed. The P value of each gene signaling pathway target is presented in a negative value of logarithmic form (-lgP), which indicate that the higher the value, the higher the significance level of the gene signaling pathway."

Figure 4

Global signal transduction network constructed with potential gene targets of miR-106b-5p and the upstream and downstream genes in human umbilical vein endothelial cells Using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, the relationship between the miR-106b-5p-targeting genes and the upstream and downstream genes/proteins could be found, and the global signal transduction network (interaction network) between genes could then be constructed. Red spots represented upregulated genes, and blue spots indicated downregulated genes."

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