Expression of miR-106b-5p in children with primary immune thrombocytopenia and its correlation with T cells / 中国当代儿科杂志

OBJECTIVES@#To study the expression level of plasma miR-106b-5p in primary immune thrombocytopenia (ITP) and its correlation with the levels of T helper 17 cell (Th17) and regulatory T cell (Treg) and the Th17/Treg ratio.@*METHODS@#A total of 79 children with ITP (ITP group) and 40 healthy children (control group) were selected as subjects. According to the treatment response, the 79 children with ITP were divided into three groups: complete response (n=40), partial response (n=18), and non-response (n=21). Quantitative real-time PCR was used to measure the expression level of miR-106b-5p. Flow cytometry was used to measure the frequencies of Th17 and Treg, and the Th17/Treg ratio was calculated. The correlation of the expression level of plasma miR-106b-5p with the frequencies of Th17 and Treg and the Th17/Treg ratio was analyzed.@*RESULTS@#Compared with the control group, the ITP group had significantly higher levels of miR-106b-5p, Th17, and Th17/Treg ratio (P<0.05) and a significantly lower level of Treg (P<0.05). After treatment, the ITP group had significant reductions in the levels of miR-106b-5p, Th17, and Th17/Treg ratio (P<0.05) and a significant increase in the level of Treg (P<0.05). Compared with the partial response and non-response groups, the complete response group had significantly lower levels of miR-106b-5p, Th17, and Th17/Treg ratio (P<0.05) and a significantly higher level of Treg (P<0.05). The correlation analysis showed that in the children with ITP, the expression level of plasma miR-106b-5p was positively correlated with the Th17 level and the Th17/Treg ratio (r=0.730 and 0.816 respectively; P<0.001) and was negatively correlated with the Treg level (r=-0.774, P<0.001).@*CONCLUSIONS@#A higher expression level of miR-106b-5p and Th17/Treg imbalance may be observed in children with ITP. The measurement of miR-106b-5p, Th17, Treg, and Th17/Treg ratio during treatment may be useful to the evaluation of treatment outcome in children with ITP.

BRD4 inhibitor specifically inhibits the development of wild-type Kras differentiated thyroid carcinoma by regulating BRD4/miR-106b-5p/P21 axis / 国际肿瘤学杂志

Objective:To explore the influence of bromodomain-containing protein 4 (BRD4) inhibitor on wild-type Kras differentiated thyroid carcinoma (DTC) and its mechanism.Methods:The DTC cell line Kras WT TPC-1 was selected and the mutant Kras G12D TPC-1 cells were constructed. CCK-8 assay was used to detect the effect of BRD4 inhibitor JQ-1 on the proliferation activity of Kras WT TPC-1 cells. Kras WT TPC-1 cells were treated with 0.2 μmol/L JQ-1 (JQ-1 group), and a negative control group (NC group) was set. Transwell invasion assay and flow cytometry were used to detect the effect of JQ-1 on the invasion and apoptosis of Kras WT TPC-1 cells. The effect of JQ-1 on the expressions of BRD4, miR-106b-5p and P21, and the effect of P21 inhibitor UC2288 on the expressions of P21 and BRD4 were detected. Kras WT TPC-1 cells were divided into JQ-1+ NC-OE group, JQ-1+ p21-OE group (overexpression of p21) and JQ-1+ p21-OE+ miR-106b-5p mimic group (overexpression of p21 and miR-106b-5 at the same time), and the proliferation, invasion and apoptosis of cells in each group were detected. TPC-1 cells were divided into Kras WT group, Kras WT+ JQ-1 group, Kras G12D group and Kras G12D+ JQ-1 group, and the cell proliferation, invasion and apoptosis of each group were detected. Results:JQ-1 inhibited the proliferation activity of Kras WT TPC-1 cells in a dose-dependent and time-dependent manner. In the NC group and JQ-1 group, the numbers of cell invasion were 124.67±9.61 and 82.67±8.02, and the apoptosis rates were (5.91±0.34)% and (10.33±1.10)%, respectively, with statistically significant differences ( t=5.812, P=0.004; t=6.653, P=0.003). JQ-1 significantly inhibited the expressions of BRD4 and miR-106b-5p, and promoted the expression of P21 in Kras WT TPC-1 cells. UC2288 significantly inhibited P21 expression, but had no significant effect on BRD4 expression. In the JQ-1+ NC-OE group, JQ-1+ p21-OE group and JQ-1+ p21-OE+ miR-106b-5p mimic group, the proliferation activities at 24 h of Kras WT TPC-1 cells was 0.46±0.03, 0.35±0.04 and 0.44±0.03 ( F=8.720, P=0.017), and the proliferation activity of JQ-1+ p21-OE group was significantly lower than that of the JQ-1+ NC-OE group ( P<0.05). The numbers of cell invasion in the three groups were 83.00±9.17, 56.67±6.03 and 79.67±10.07 ( F=8.347, P=0.018), and the number of cell invasion in the JQ-1+ p21-OE group was significantly lower than that in the JQ-1+ NC-OE group ( P=0.009). The apoptosis rates of the three groups were (10.00±0.49)%, (15.39±1.14)% and (10.32±0.80)% ( F=37.764, P<0.001), and the apoptosis rate of the JQ-1+ p21-OE group was significantly higher than that in the JQ-1+ NC-OE group ( P<0.001). There were no significant differences in cell proliferation activity, invasion number and apoptosis rate between JQ-1+ p21-OE+ miR-106b-5p mimic group and JQ-1+ NC-OE group (all P>0.05). In Kras WT group, Kras WT+ JQ-1 group, Kras G12D group and Kras G12D+ JQ-1 group, the cell proliferation activities at 24 h were 0.50±0.05, 0.39±0.04, 0.68±0.08 and 0.64±0.05 ( F=17.776, P<0.001). Compared with the Kras WT group, cell proliferation activity in the Kras WT+ JQ-1 group was significantly decreased, while that in the Kras G12D group was significantly increased (both P<0.05). The numbers of cell invasion in the four groups were 129.33±11.50, 86.00±9.54, 161.67±13.01 and 146.33±13.20 ( F=22.598, P<0.001). Compared with the Kras WT group, the number of cell invasion in the Kras WT+ JQ-1 group was significantly decreased ( P=0.002), and that in the Kras G12D group was significantly increased ( P=0.010). The apoptosis rates in the four groups were (6.17±0.50)%, (10.42±0.73)%, (3.43±0.47)% and (3.41±0.32)% ( F=119.170, P<0.001). Compared with the Kras WT group, the apoptosis rate in the Kras WT+ JQ-1 group was significantly increased ( P<0.001), and that in the Kras G12D group was significantly decreased ( P<0.001). There were no significant differences in cell proliferation activity, invasion number and apoptosis rate between Kras G12D+ JQ-1 group and Kras G12D group (all P>0.05). Conclusion:BRD4 inhibitor can specifically inhibit the development of wild-type Kras DTC via regulating the molecular axis of BRD4/miR-106b-5p/P21, but has no significant effect on the proliferation, invasion and apoptosis of mutant Kras DTC tumor cells.

Up-regulated miR-106b inhibits ox-LDL-induced endothelial cell apoptosis in atherosclerosis

This research aimed to explore the molecular mechanism of microRNA (miR)-106b in cell apoptosis of atherosclerosis (AS). Human aortic endothelial cells (HAECs) were divided into control group, oxidized-low-density lipoproteins (ox-LDL) group, miR-106b NC+ox-LDL group, miR-106b mimics+ox-LDL group, miR-106b mimics+PTEN+ox-LDL group, and miR-106b mimics+empty+ox-LDL group. Real-time fluorescence quantitative polymerase chain reaction, cholecystokinin, TdT-mediated biotinylated nick end-labeling assay, luciferase reporter gene assay, and flow cytometry analysis were performed to determine the morphology, proliferation, and apoptosis in HSECs. Moreover, the levels of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), Bcl-2, p-P13K, and p-AKT in HAECs were detected by western blot. MiR-106b was down-regulated in ox-LDL-induced HAECs. PTEN was the target gene of miR-106b-5p. Overexpression of PTEN inhibited the anti-apoptotic effect of miR-106b. Compared with the control group, the proportion and number of HAECs apoptosis and Bax, caspase-3, and caspase-9 expression in ox-LDL and miR-106b mimics+PTEN+ox-LDL groups were significantly increased (all P<0.05). Moreover, the activity of HAECs and Bcl-2 were decreased significantly (all P<0.05). Overexpression of miR-106b in ox-LDL-induced AS inhibited endothelial cell apoptosis. Furthermore, miR-106b might activate the PI3K/AKT pathway by down-regulating the expression of PTEN in ox-LDL-induced HAECs.

miR-106b induces the apoptosis and inhibits the proliferation of nasopharyngeal carcinoma cells / 西安交通大学学报(医学版)

Objective To investigate the effect of miR-106b on the apoptosis and proliferation of nasopharyngeal carcinoma (NPC ) cells. Methods We analyzed differences in miRNA expression in nasopharyngeal carcinoma and adjacent normal tissues with miRNA microarray.Taq Man miRNA detection kit and Real-time fluorescence quantitative PCR were used to detect the expressions of miR-106 and RhoC mRNA in nasopharyngeal carcinoma and adjacent tissues.The miR-106b and target gene binding sites were predicted with miRnada.The target gene was verified by double luciferase.Western blot was used to detect the expression of RhoC regulated by miR-106b.Annexin and TUNEL were used to detect the effect of miR-106b on the apoptosis of nasopharyngeal carcinoma cells;the effect of miR-106b on the proliferation of nasopharyngeal carcinoma cells was detected by MTT assay.Results miRNA microarray analysis showed that the expression of miR-106b was lower in NPC tissues than in adjacent normal tissues.The results of RT-PCR showed that the expression of miR-106b in nasopharyngeal carcinoma was decreased (P <0.05)while the expression of RhoC was increased in nasopharyngeal carcinoma (P <0.05).The expressions of miR-106b and RhoC in NPC were negatively correlated (r =-0.5866, P <0.001).The results of luciferase reporter assay showed that the activity of luciferase in miR-106b group was lower than that in empty plasmid group (P < 0.05 ).The results of Western blot showed that miR-106b could decrease the expression of RhoC in NPC tissues (P <0.05).Annexin V-PI and TUNEL showed that the apoptosis ofnasopharyngeal carcinoma cells was significantly higher in miR-106 group than in empty plasmid group (P <0.05). MTT results showed that the proliferation of nasopharyngeal carcinoma cells in miR-106b group was lower than that in empty plasmid group (P <0.05).Conclusion miR-106b may induce the apoptosis of nasopharyngeal carcinoma cells and inhibit the proliferation of nasopharyngeal carcinoma cells by down-regulating the expression of RhoC.

miR-106b induces the apoptosis and inhibits the proliferation of nasopharyngeal carcinoma cells / 西安交通大学学报(医学版)

Objective To investigate the effect of miR-106b on the apoptosis and proliferation of nasopharyngeal carcinoma (NPC ) cells. Methods We analyzed differences in miRNA expression in nasopharyngeal carcinoma and adjacent normal tissues with miRNA microarray.Taq Man miRNA detection kit and Real-time fluorescence quantitative PCR were used to detect the expressions of miR-106 and RhoC mRNA in nasopharyngeal carcinoma and adjacent tissues.The miR-106b and target gene binding sites were predicted with miRnada.The target gene was verified by double luciferase.Western blot was used to detect the expression of RhoC regulated by miR-106b.Annexin and TUNEL were used to detect the effect of miR-106b on the apoptosis of nasopharyngeal carcinoma cells;the effect of miR-106b on the proliferation of nasopharyngeal carcinoma cells was detected by MTT assay.Results miRNA microarray analysis showed that the expression of miR-106b was lower in NPC tissues than in adjacent normal tissues.The results of RT-PCR showed that the expression of miR-106b in nasopharyngeal carcinoma was decreased (P <0.05)while the expression of RhoC was increased in nasopharyngeal carcinoma (P <0.05).The expressions of miR-106b and RhoC in NPC were negatively correlated (r =-0.5866, P <0.001).The results of luciferase reporter assay showed that the activity of luciferase in miR-106b group was lower than that in empty plasmid group (P < 0.05 ).The results of Western blot showed that miR-106b could decrease the expression of RhoC in NPC tissues (P <0.05).Annexin V-PI and TUNEL showed that the apoptosis ofnasopharyngeal carcinoma cells was significantly higher in miR-106 group than in empty plasmid group (P <0.05). MTT results showed that the proliferation of nasopharyngeal carcinoma cells in miR-106b group was lower than that in empty plasmid group (P <0.05).Conclusion miR-106b may induce the apoptosis of nasopharyngeal carcinoma cells and inhibit the proliferation of nasopharyngeal carcinoma cells by down-regulating the expression of RhoC.

MiR-106b activate Wnt/β-catenin pathway in HCC cells / 实用医学杂志

Objective To investigate the influence of miR-106b on Wnt/β-catenin pathway in HCC cells. Methods QGY-7703 and HepG2 cells were transfected with miRNA mimics or inhibitors. TOP/FOP luciferase ratio assay was used to test the Wnt/β-catenin pathway activity. The expression of downstream targeted genes of Wnt/β-catenin pathway were examined by Real-time PCR. The accumulation of β-catenin in nuclears were measured by Western blotting. Results Ectopic expression of miR-106b dramatically increased the average TOP/FOP ratio and the mRNA expression of downstream targeted genes in QGY-7703 and HepG2 cells. Compared with that in control cells , miR-106b over-expression promoted the nuclear β-catenin accumulation in QGY-7703 cells. Clonclusion MiR-106b activated Wnt/β-catenin pathway in HCC cells.

Target genes of the hsa-miR-106b-25 cluster:Prediction and bioinformatic analysis / 医学研究生学报

Objective The hsa-miR-106b-25 gene cluster is involved in various biological processes of carcinoma .This study aims at a prediction and function analysis of the hsa-miR-106b-25 gene cluster, miR-106b, miR-93, and miR-25, so as to provide some evidence for further studies on the functions of the three miRNAs and the mechanisms of their interaction . Methods We obtained the sequences of miR-106b, miR-93, and miR-25 from the miRBase, predicted their target genes with TargetScan , PicTar, and miRanda, and used 3 or more experimentally verified target genes from the miRTarbase as the gene set for further bioinformatic analysis .We predic-ted the biological processes of the target genes by GeneOntology analysis and enriched KEGG ( Kyoto Encyclopedia of Genes and Genomes) by pathway analysis, produced protein-protein interaction ( PPI) networks with STRING . Results The target genes of the miR-106b-25 gene cluster were significantly enriched in such biological processes as the regulation of macromolecule metabolism , regulation of metabolic process , and cell cycle process , while the KEGG pathway mainly in glioma, melanoma, prostate cancer , and gallbladder carcino-ma.The proteins encoded by the targeted genes of showed complicated interactions , and those encoded by the KAT2B, PTEN, TP53, CDH1, MDM2, E2F1, RB1, and SMAD7 plaid a core role in the interac-tion network. Conc lusoi n MiRNAs of the miR-106b-25 gene cluster regulate the downstream target proteins involved in tumorigenesis by participating in the cell cycle and cancer signaling pathway .

A study on the expression of miR-106b~25 cluster in human glioma / 中国肿瘤临床

Objective:To detect the expression of miR-106b~25 cluster in glioma cell line and tissues. Methods:Real-time PCR was performed to determine the expression of miR-106b~25 cluster members (miR-106b, miR-93, and miR-25) in different human glio-blastoma cell lines. Different pathological grade glioma specimens were surgically removed. In-situ hybridization was performed to de-tect the expression of miR-106b~25 cluster members in different pathological levels of glioma tissues. Results:In the expression of the benchmark on normal brain tissues, three kinds of miRNAs in all test cell lines have a tendency to increase. Based on the expression of the pathological level I average rate in 43 cases of glioma specimens collected after neurosurgical operations, the real-time PCR results showed that the average expression quantity of the three kinds of miRNAs in each group gradually increase. The increase in tumor path-ological levels results in statistically significant expression differences of miR-106b and miR-93 between the groups (F=4.479, P=0.018 and F=3.493, P=0.040, respectively). However, miR-25 expression differences between the groups have no statistically signifi-cant differences (F=2.766, P=0.075). In situ hybridization results show that the expressions of three miRNAs in high grade gliomas are significantly higher than that in the low-level tumor. Spearman rank correlation analysis results indicate that the expression of these miRNAs signal-intensity distribution is positively correlated with glioma, in accordance with WHO pathology classification. The corre-lation coefficient for miR-106b, miR-93, and miR-25 are 0.617, 0.438, and 0.463, respectively (P<0.001). Conclusion:The expression of miR-106b~25 cluster members is up-regulated in the glioma and is positively correlated with tumor grade.

Advances in study of miR-106b-25 cluster in cancer / 国际肿瘤学杂志

miR-106b-25 cluster is composed of miR-106b,miR-93 and miR-25,and is a paralogue of miR-17-92 cluster.Some studies have shown that the members in miR-106b-25 cluster abundantly expressed in many cancers.Over-expressions of these miRNAs promote the growth of tumor cells by negatively regulating p21 and p57,and suppress the apoptosis of tumor cells through inhibition of Bim.Moreover,high expression of miR-106b-25 cluster might endue tumor cells with resistance to inhibitory effect of cell growth induced by TGF-β signaling.