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Long noncoding RNAs (lncRNAs) play a critical role in the regulation of atherosclerosis. Here, we investigated the role of the lncRNA growth arrest-specific 5 (lncR-GAS5) in atherogenesis. We found that the enforced expression of lncR-GAS5 contributed to the development of atherosclerosis, which presented as increased plaque size and reduced collagen content. Moreover, impaired autophagy was observed, as shown by a decreased LC3II/LC3I protein ratio and an elevated P62 level in lncR-GAS5-overexpressing human aortic endothelial cells. By contrast, lncR-GAS5 knockdown promoted autophagy. Moreover, serine/arginine-rich splicing factor 10 (SRSF10) knockdown increased the LC3II/LC3I ratio and decreased the P62 level, thus enhancing the formation of autophagic vacuoles, autolysosomes, and autophagosomes. Mechanistically, lncR-GAS5 regulated the downstream splicing factor SRSF10 to impair autophagy in the endothelium, which was reversed by the knockdown of SRSF10. Further results revealed that overexpression of the lncR-GAS5-targeted gene miR-193-5p promoted autophagy and autophagic vacuole accumulation by repressing its direct target gene, SRSF10. Notably, miR-193-5p overexpression decreased plaque size and increased collagen content. Altogether, these findings demonstrate that lncR-GAS5 partially contributes to atherogenesis and plaque instability by impairing endothelial autophagy. In conclusion, lncR-GAS5 overexpression arrested endothelial autophagy through the miR-193-5p/SRSF10 signaling pathway. Thus, miR-193-5p/SRSF10 may serve as a novel treatment target for atherosclerosis.
Asunto(s)
Humanos , Aterosclerosis/genética , Autofagia/genética , Proteínas de Ciclo Celular/metabolismo , Células Endoteliales/metabolismo , Endotelio/metabolismo , MicroARNs/metabolismo , Proteínas Represoras/metabolismo , Factores de Empalme de ARN , Factores de Empalme Serina-Arginina/genética , ARN Largo no Codificante/metabolismoRESUMEN
Objective:To investigate the expression and clinical significance of serum miR-193-5p and miR-196-5p in children with IgA nephropathy (IgAN).Methods:The 95 children with IgAN (IgAN group), 80 children with non IgAN nephritis (non IgAN group) and 50 normal subjects (control group) in Sanya Women and Children's Hospital of Shanghai Children's Medical Center and Sanya People's Hospital were selected to be included in this study. According to Oxford classification score (MEST), children with IgAN were divided into 35 cases with MEST≥3 points and 60 cases with MEST<3 points. The serum levels of miR-193-5p, miR-196-5p, IgA/C3 and galactose deficient IgA1 molecule (Gd-IgA1) in each group were compared. The receiver operating characteristic (ROC) curve was drawn to analyze the value of miR-193-5p, miR-196-5p, IgA/C3 and Gd-IgA1 in the diagnosis of IgAN. Pearson correlation analysis was used to analyze the correlation between the expression levels of miR-193-5p, miR-196-5p and IgA/C3 and Gd-IgA1.Results:The serum levels of miR-193-5p, miR-196-5p, IgA/C3 and Gd-IgA1 in IgAN group were significantly higher than those in non IgAN group and control group (all P<0.001). The levels of miR-193-5p, miR-196-5p, IgA/C3 and Gd-IgA1 in MEST≥3 group were significantly higher than those in MEST<3 group (all P<0.001). ROC curve analysis showed that the area under the curve for the combined diagnosis of IgAN by miR-193-5p, miR-196-5p, IgA/C3 and Gd-IgA1 was 0.958 (95% CI: 0.902-0.998), with sensitivity of 98.6% and specificity of 86.4%. Pearson correlation analysis showed that the expression levels of serum miR-193-5p and miR-196-5p in IgAN children were positively correlated with IgA/C3 and Gd-IgA1 (all P<0.001). Conclusions:The expression levels of serum miR-193-5p and miR-196-5p were significantly increased in children with IgAN, and the combined detection of IgA/C3 and Gd-IgA1 has high value for the diagnosis of IgAN in children.
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@#[Abstract] Objective: To investigate the effect of lncRNA SNHG11 on the proliferation, invasion and migration of non-small cell lung cancer (NSCLC) A549 cells and its possible mechanisms. Methods: qPCR was used to detect the levels of lncRNA SNHG11 and miR-193a-5p in human embryonic lung cells (HEL-1) and lung cancer cells (A549, H1299, and HCC827). A549 cells were transfected with SNHG11 small interfering RNA (si-SNHG11), miR-193a mimic or miR-193a inhibitor. The proliferation of A549 cells was detected by CCK-8 assay, migration and invasion of A549 cells were detected by Wound healing and Transwell assay, the protein expression of Ki67 and Cyclin D1 was determined by Western blot, and the targeting relationship between lncRNA SNHG11 and miR-193a-5p was verified by Dual-luciferase reporter experiment. Results: Compared with HEL-1 cells, the expression level of lncRNA SNHG11 was significantly increased while the expression of miR-193a-5p was decreased in lung cancer A549, H1299 and HCC827 cells (all P<0.05). Silencing lncRNA SNHG11 inhibited the proliferation, migration and invasion of A549 cells and reduced the protein expression of Ki67 and Cyclin D1 (all P<0.05). Over-expression of miR-193a-5p inhibited the proliferation, migration and invasion of A549 cells (all P<0.05). lncRNA SNHG11 could targetedly adsorb miR-193a-5p. miR-139a-5p inhibition could partially reverse the effect of silencing lncRNA SNHG11 on the proliferation, invasion and migration of A549 cells (all P<0.05). Conclusion: lncRNA SNHG11 promotes the proliferation, invasion and migration of NSCLC cells by adsorbing miR-193a-5p.
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Reportedly, circular RNAs (circRNAs) are crucial regulators in cancer progression. Nonetheless, the molecular mechanism of circRNAs in hepatocellular carcinoma (HCC) has not been fully clarified. Gene expression omnibus (GEO) database was employed to screen out the differentially expressed circRNAs in HCC. qRT-PCR and western blot were executed to detect circ_0001806 expression, miR-193a-5p expression, and MMP16 mRNA and protein expressions in HCC. The effect of circ_0001806 on HCC was analyzed by the CCK-8 method and Transwell experiment. RIP assay, pull-down experiment, and dual-luciferase reporter gene experiment were applied to validate the targeting relationships among circ_0001806, miR-193a-5p, and MMP16. Circ_0001806 was up-modulated in HCC tissues and cell lines. Knockdown of circ_0001806 impeded the multiplication, migration, and invasion of HCC cells. Circ_0001806 could up-regulate MMP16 expression through repressing miR-193a-5p, thereby facilitating the malignant biological behaviors of HCC. Circ_0001806 promoted HCC progression by regulating miR-193a-5p/MMP16 axis.
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Objective To investigate the expression level of serum miR-193a-3p,miR-337-5p and miR-483-5p in esophageal squamous cell carcinoma (ESCC) patients and to explore their value for diagnosis and prognosis of ESCC.Methods Serum samples were collected from 63 ESCC patients before and after surgery in Nanjing General Hospital and Xuzhou Cancer Hospital between June 2013 and May 2014 and serum sanples of 63 age-and sex-matched healthy individuals acted as the normal controls.TaqMan Low Density Assay was used to detect the deregulated miRNA in ESCC patients and then quantitative real-time PCR was used to validate the upregulated miRNA miR-193a-3p,miR-337-5p and previously reported miR483-5p that was upregulated in oral squamous cell carcinoma (OSCC).Finally,the three miRNA were evaluated for their clinical value in the diagnosis and predicting prognosis of ESCC.Results Compared with normal controls,serum levels of miR193a-3p,miR-337-5p and miR-483-5p in ESCC patients were significantly up-regulated (0.459±0.339 vs 0.195±0.084,U =591;5.686±5.211 vs 2.476±0.808,U=605;32.545 ± 22.479 vs 19.509±10.601,U=1 037,respectively,all P＜ 0.0001) and their levels were significantly reduced after the surgical treatment (P＜0.05).The areas under the ROC curve of serum miR-193a-3p,miR-337-5p,miR-483-5p and miR-Panel were all larger than that of CEA.Univariate Kaplan-Meier analysis revealed that ESCC patients with low expression level of miR-483-5p in postoperative serum exhibited higher survival rate than those with high level(P=0.022).Conclusion Serum miR-193a-3p,miR-337-5p and miR-483-5p can be potential molecular biomarkers in the diagnosis and predicting prognosis of ESCC.
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Objective To investigate the significance of miR-193b to biological behaviors of hepatocellular carcinoma (HCC). Methods 48 cases of HCC specimens and corresponding adjacent tissues were collected, and the miR-193b expression levels in these specimens were measured by real-time quantitative PCR. The miR-193b expression was measured by the same way in HepG2 and SMMC-7721 cells. The HepG2 and SMMC-7721 were transfected with miR-193b mimics or negative control miRNA mimic with Lipofectamine 2000, and the non-transfected cells were taken as blank control. The proliferation ability of the HCC cells were detected by MTT method, and the apoptosis rate was tested by flow cytometry. Results The expression level of miR-193b in HCC tissues (2.441 ±0.569) was significantly lower than that in the corresponding adjacent tissues (15.488±4.326) (P < 0.05). Compared with normal liver cell line L-O2, the expression levels of miR-193b were significantly lower in HepG2 and SMMC-7721 cells. Transfected with miR-193b mimic, the proliferation ability of HepG2 and SMMC-7721 cells were reduced, while their apoptosis were increased. Conclusion miR-193b may be negative to regulate the proliferation of HCC and increase its apoptosis.
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Objective To investigate the role of miR?193a?3p in the radioresistance of esophageal squamous cell carcinoma ( ESCC) . Methods MTT assay was used to identify the cell lines with the highest radiosensitivity and radioresistance in four esophageal cancer cell lines exposed to irradiation of 6 MV X?ray. Stem?loop quantitative real?time PCR was used to measure the expression levels of miR?193a?3p, miR?155, and miR?22?3P in the two cell lines. Further studies were performed on miR?193a?3p because of the substantial difference in its expression between the two cell lines. The mimic (3PM) and antagomiR (3PA) of miR?193a?3p as well as siRNA ( si?LOXL4) were synthesized and transfected into cells to elevate and inhibit miR?193a?3p expression. MTT assay and flow cytometry were used to evaluate the effects of miR?193a?3p and its downstream gene LOXL4 on radiosensitivity. Results KYSE510 and KYSE410 were characterized as cell lines with the highest radiosensitivity and radioresistance, respectively. miR?193a?3p had a substantially larger difference in expression between the two cell lines than miR?155 or miR?22?3P (1. 00 ∶ 21. 00). Transfection of 3PM resulted in elevated expression of miR?193a?3p in KYSE510, which had a significantly lower radiosensitivity and a significantly reduced apoptosis ratio by 11. 01% compared with the control group ( P<0. 05) . KYSE410 transfected with 3PA had a significantly higher radiosensitivity ( P<0. 05) . The expression of LOXL4, a downstream gene of miR?193a?3p, was negatively correlated with miR?193a?3p expression. Transfection with si?LOXL4 inhibited the expression of LOXL4, which resulted in a significantly lower radiosensitivity and a significantly reduced apoptosis ratio by 7. 07% compared with the control group ( P<0. 05) . Conclusions miR?193a?3p promotes the radioresistance of esophageal cancer cells probably by regulation of LOXL4.
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Objective To explore the value of the plasma miR-193a-5p level on diagnosis and monitoring the response to treatment in acute myeloid leukemia (AML). Methods Peripheral blood samples were obtained from AML patients enrolled in hematology department of the Second Hospital of Shanxi Medical University from July 2015 to December 2015, including 30 de novo AML patients, 9 patients in complete remission (CR) and 6 patients in relapse. Peripheral blood samples from 15 healthy people were randomly choosed as the health control group. Plasma miR-193a-5p expression levels were detected by using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Results The plasma miR-193a-5p relative expression level of AML patients group [0.465 6 (0.103 1-5.000 2)] was obviously lower than that of health control group [0.766 1 (0.052 1-3.134 4)] (U= 122, P= 0.018 7). The plasma miR-193a-5p relative expression levels of de novo group and relapse AML group were significantly lower than those of CR group and health control group (P<0.05), and there was no significant difference between the CR group and health control group (U= 56, P= 0.511 9). No significant correlation was found between the plasma miR-193a-5p level and age, gender, blast percentage of the bone marrow, peripheral blood leukocyte count, platelet count, CD34+cells'percentage and so on. Conclusion The decreased plasma miR-193a-5p expression level can be served as a new and noninvasive biomarker for the evaluation of diagnosis and treatment in AML.
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Objective To determine the levels of miR-193a-3p in serum from patients with renal clear cell carcinoma,and eval-uate the potential of miR-193a-3p asa molecular biomarker of renal clear cell carcinoma.Methods Serum samples were taken from 107 untreated hospitalized patients with renal clear cell carcinoma (TNM Ⅰ grade 76 cases,Ⅱ grade 16 cases,Ⅲ grade 2 cases,Ⅳ grade 8 cases and unknown 5 cases)in Nanjing General Hospital from 2010 to 2014 year and 107 age-and gender-matched healthy individuals to determine the level of miR-193a-3p by quantitative reverse-transcription PCR (qRT-PCR). The early diagnostic value of miR-193a-3p for renal clear cell carcinoma was assessed by ROC curve.Results qRT-PCR confirmed that serum miR-193a-3p expression levels was significantly elevated in cancer than in normal controls (3.294± 1.526 vs 1.944±0.600,P =0.000).TNM Ⅰ grade (3.411±1.676 vs 1.944±0.600,P =0.000),Ⅱ grade (2.926±0.927 vs 1.944±0.600,P =0.001),Ⅳ grade(2.926±0.894 vs 1.944±0.600,P =0.000)is significantly higher than that in con-trol group.The area under the ROC curve (AUC)of miR-193a-3p was 0.820 (95% CI,0.756 ~ 0.883),demonstratinga high sensitivity (80.3%)and specificity (93.5%)for the early diagnosis of renal clear cell carcinoma.Conclusion MiR-193a-3p content was significantly elevated in serum from renal clear cell carcinoma and has a high accuracy in diagnosing ear-ly cancer,which holds potential as molecular biomarker for renal clear cell carcinoma.
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AIM:To investigate the effects of microRNA-193 (miR-193) on the proliferation and apoptosis of rat bone marrow mesenchymal stem cells (MSCs).METHODS:Cultured rat MSCs were transfected with pre-miR-193 or anti-miR-193 to regulate the expression of miR-193.The proliferation of the MSCs after transfection was evaluated by MTS assay, colorimetric BrdU cell proliferation assay and Ki-67 immunostaining.Cell apoptosis was analyzed by flow cytometry with Annexin V/PI staining.The effect of miR-193 on the expression of cell cycle-related proteins was evaluated by qRT-PCR.RESULTS:Transfection of pre-miR-193 or anti-miR-193 regulated the expression of miR-193 in MSCs effectively . Over-expression of miR-193 significantly promoted the proliferation of MSCs ( P0.05).The result of qRT-PCR indicated miR-193 promoted the expression of cyclin-dependent kinase 2 ( CDK2 ) significantly ( P<0.01).CONCLUSION:miR-193 promotes the proliferation of MSCs possibly through the CDK 2 pathway.
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Objective To analyze the expression and regulation of HES1 in sperms with low motility.Methods Thirty semen samples from asthenospermia patients and 20 semen samples from healthy and fertile adults were collected,total RNAs were extracted to produce cDNAs probes.Hybridization with Phalanx OneArray~(TM) containing 30 968 probes was carried out after the labeled cDNAs were purified by PCR product purification kit.Realtime RT-PCR was used to analyze the expression of hsa-miR-487a and hsa-miR-193b;the expression of the target genes of hsa-miR-487a and hsa-miR-193b were searched from gene-expression profiles in asthenospermia patients' sperms.Results The expression level of HES1 in low motility sperms was up-regulated.The expression level of hsamiR-193b in low motility sperms was 2.19 times higher than that in high motility sperms,hsa-miR-487a was 0.43% of that in high motility sperms.Conclusion The expression level of HES1 in low motility sperms was up-regulated.Hsa-miR-487a and hsa-miR-193b may affect the expression of HES1 and so regulate sperm motility.