ABSTRACT
Objective:To explore and analyze the value of detection of peripheral blood miR-194 combined with fecal miR-143 in the clinical screening of colorectal cancer.Methods:A total of 83 patients diagnosed with colorectal cancer by pathological tissue admitted to Huangshi Hospital of Traditional Chinese Medicine of Hubei Province from October 2019 to October 2020 were selected as the observation group, and 50 healthy volunteers who underwent physical examinations during the same period were selected as the control group. The levels of miR-194 in peripheral blood and miR-143 in feces were detected by fluorescence quantitative PCR. The level difference between the two groups and their correlations with clinicopathological parameters of patients with colorectal cancer were analyzed. Receiver operating characteristic (ROC) curve was drawn based on peripheral blood miR-194 and fecal miR-143 to evaluate their value for clinical screening of colorectal cancer.Results:The level of miR-194 in peripheral blood of the observation group was significantly higher than that of the control group (1.91±0.34 vs. 0.76±0.23) , while the level of fecal miR-143 in the observation group being significantly lower than that of the control group (1.85±0.43 vs. 2.48±0.62) , with statistically significant differences ( t=21.16, P<0.001; t=6.91, P<0.001) . Age of patients with colorectal cancer ( t=0.83, P=0.408; t=1.17, P=0.244) , TNM stage ( t=1.03, P=0.307; t=0.11, P=0.909) , lymphatic metastasis ( t=0.37, P=0.711; t=1.85, P=0.068) , distant metastasis ( t=0.41, P=0.683; t=1.72, P=0.089) were not correlated with the levels of peripheral blood miR-194 and fecal miR-143. When the cut-off value of miR-194 in peripheral blood was 1.82, the area under the ROC curve for the diagnosis of colorectal cancer was 0.76, and the diagnostic sensitivity and specificity were 79.38% and 74.29%, respectively. When the cut-off value of fecal miR-143 was 2.16, the area under the ROC curve for the diagnosis of colorectal cancer was 0.71. At this time, the diagnostic sensitivity and specificity were 76.54% and 73.61%, respectively. The area under ROC curve of combined detection for colorectal cancer was 0.81, and the diagnostic sensitivity and specificity were 83.46% and 75.43%, respectively. Conclusion:Peripheral blood miR-194 is highly expressed in colorectal cancer patients, and fecal miR-143 is low in colorectal cancer patients. The combined detection of the two has a high sensitivity for early diagnosis of colorectal cancer, which can provide important reference basis for early diagnosis of colorectal cancer and has high clinical application value.
ABSTRACT
Objective: To detect the expression levels of miR-194-5p and magnesium transporter protein 1 (MagTl) in peripheral blood mononuclear cells (PBMCs) of the patients with chronic hepatitis B virus (HBV) infection, and to investigate their possible clinical significances. Methods: The clinical materials of 40 healthy subjects (healthy control group). 40 cases of asymptomatic hepatitis B carriers (HBV carrier group). 40 patients with mild-moderate chronic hepatitis B (mild-moderate CHB group) and 40 patients with severe chronic hepatitis B (severe CHB group) were collected. The peripheral blood of the subjects in various groups was collected and the PBMCs were isolated by Ficoll-Hypaque density gradient centrifugation. The expression levels of miR-194-5p and MagTl mRNA in PBMCs of the subjects in various groups were detected by RT-PCR assay, the expression levels of MagTl protein in PBMCs of the subjects in various groups were detected by Western blotting method, the levels of Mg in peripheral blood and PBMCs of the subjects in various groups were detected, and the expression levels of CD6+ T lymphocyte surface molecules programmed death receptor 1 (PD-1) and natural killer cell receptor 2 group D molecule (NKG2D) in PBMCs of the subjects in various groups were analyzed by flow cytometry. The miR-194-5p mimic (miR-194-5p mimic group) and miR-jNC (miR-NC group) were transfected into the 293T cells, and then the dual luciferase reporter gene assay was used to detect the luciferase activities in 293T cells in two groups. Results: Compared with healthy control group, the expression levels of MagTl mRNA and protein, the expression levels of CD8 + T lymphocyte surface molecule NKG2D and the levels of Mg in PBMCs of the patients in HBV carrier group, mild-moderate CHB group and severe CHB group were decreased ( P<-0. 05) . the expression levels of miR-194-5p mRNA in PBMCs and CD8 + T lymphocyte surface molecule PD-1 of the patients were increased (P<.0. 05). Compared with HBV carrier group and mild-moderate CHB group, the expression levels of MagTl mRNA and protein, the expression level of CD8+ T lymphocyte surface molecule NKG2D and the level of Mg in PBMCs of the patients in severe CHB group were decreased ( P<0. 05). and the expression levels of miR-194-5p mRNA in PBMCs and CD8 + T lymphocyte surface molecule PD-1 of the patients were increased (P<0. 05). Compared with HBV carrier group, the expression levels of MagTl mRNA and protein, the expression level of CD8+ T lymphocyte surface molecule NKG2D and the level of Mg in PBMCs of the patients in mild-moderate CHB group were decreased ( P
ABSTRACT
Objective@#To investigate the levels of urinary exosomal miR-194-5p in children with idiopathic nephrotic syndrome (INS) and its clinical value as a non-invasive auxiliary diagnostic marker. @*Methods@#Urine samples were collected from 101 INS children and 98 sex- and age-matched healthy children, and the levels of urinary exosomal miR-194-5p were determined by quantitative real-time polymerase chain reaction (qRT-PCR). The clinical value of urinary exosomal miR-194-5p in the diagnosis of children′s INS was evaluated by the ROC curve and correlation analysis. @*Results@#The levels of urinary exosomal miR-194-5p in INS children (2.420 [0.650,9.515] fmol/L) were significantly higher than that in healthy controls (0.360 [0.220,0.653] fmol/L, U=1 552, P＜0.01). Compared with the INS children before treatment, the levels of urinary exosomal miR-194-5p in clinical remission period decreased significantly (0.320 [0.145,0.523] fmol/L vs 0.975 [0.375,4.358] fmol/L, W=708, P＜0.01). Moreover, the levels of miR-194-5p in INS children with heavy urine protein (8.430 [7.225,13.070] fmol/L) were significantly higher than that with light urine protein (2.130 [1.180,3.090] fmol/L, U=0, P＜0.01). The area under the ROC curve (AUC ROC ) of miR-194-5p was 0.843 (95%CI: 0.789-0.897) for the diagnosis of INS children. Spearman correlation analysis showed that the levels of urinary exosomal miR-194-5p in INS children were negatively correlated with serum albumin levels (r=-0.300), and were positively correlated with serum total cholesterol levels (r=0.278) and 24-hour urine protein content (r=0.296, all P＜0.01). @*Conclusion@#The levels of urinary exosomal miR-194-5p in INS children increase obviously, and are closely associated with 24-hour urine protein, serum albumin and total cholesterol levels, indicating that urinary exosomal miR-194-5p may be serve as a new non-invasive fluid biopsy molecular marker for the auxiliary diagnosis of INS in children.
ABSTRACT
Objective@#To study the effect of miR-194-3p on the migration of keloid fibroblasts.@*Methods@#Differentially expressed miRNA were screened by gene chip in 8 human keloid and normal tissues. The down regulated miR-194-3p was selected for study and its binding to RUNX2 was predicted by MiRDB, and verified by fluorescent reporter gene in human keloid fibroblasts (HKFs) and passage 3 keloid cells, respectively. The effect of miR-194-3p on the migration of fibroblasts was detected by transwell assay. Western blot and real-time PCR were used to analyze the effect of miR-194-3p on RUNX2 and MMP2 expression in HKFs. The results were analyzed by SPSS 19.0 software and compared by non-paired t test. P<0.05 was considered statistically significant.@*Results@#There were abnormal expressions of miR-721, miR-21-3p, miR-382-3p, miR-194-3p, miR-3107-5p and miR-144-5p in keloid. miRDB software analysis showed that miR-194-3p and RUNX2 had targeted binding sites. Reporter gene experiments showed that miR-194-3p inhibited the activity of RUNX2 by about 50% compared with the control group (t=2.7764, P=0.0005). MiR-194-3p could significantly inhibit the expression of RUNX2 and MMP2, and inhibited the migration of keloid fibroblasts (t=2.7764, P<0.05).@*Conclusion@#MiR-194-3p may inhibit the migration of fibroblasts by inhibiting the activity of RUNX2 in keloid.
ABSTRACT
Objective:To study the expression and interaction between miR-194 and PTPN12 in the process of age-related atrophy of thymus for clarifying the regulatory mechanism in the process of this disease.Methods:C57BL/6 mouse were divided into 4 groups as 1 month,6 months,10 months and 19 months old and each group has 6 cases.Thymus tissue was removed and thymic stromal cells were isolated.And thymus epithelial cells were washed out by CD45 antibody and LS column after anesthesia.Fluorescence quantitative real-time PCR and Western blot were used to detect the changes of miR-194 and PTPN12 gene expression in thymus epithelial cells with aging.miR-194 and PTPN12 luciferase reporter vectors were transfected into HEK293 cells,and the auto fluorescence values were detected at 24 h and 48 h,respectively in vitro.Results:The expression level of miR-194 decreased (P<0.05),while the expression level of PTPN12 mRNA increased (P<0.05) as the age increased.And the correlation between miR-194 and PTPN12 mRNA expression was found to be negative(P<0.05).In vitro,luciferase reporter gene results show that miR-194 has a direct effect on the 3'UTR region of PTPN12 gene and had the highest binding efficiency in 48 h.Conclusion:PTPN12 is one of the target genes of miR-194,which is involved in the aging process of thymus and is an important factor regulating the function of thymic ep-ithelial cells.
ABSTRACT
PURPOSE: Long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) has been implicated as an oncogene in the development and progression of osteosarcoma. This study aims to explore the mechanism of NEAT1 in osteosarcoma. MATERIALS AND METHODS: Expressions of NEAT1 and miR-194 in osteosarcoma tissues and cells were detected by quantitative real-time PCR. The effects of NEAT1 knockdown or miR-194 overexpression on cell proliferation, invasion, and apoptosis were determined by 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide (MTT) assay, transwell invasive assay, and flow cytometry analysis, respectively. Luciferase reporter assay was performed to observe the possible interaction between NEAT1 and miR-194. RESULTS: NEAT1 was upregulated and miR-194 was downregulated in osteosarcoma tissues and cells. Knockdown of NEAT1 or overexpression of miR-194 suppressed proliferation and invasion and induced apoptosis of osteosarcoma cells in vitro. Luciferase reporter assay validated that NEAT1 could interact with miR-194 and negatively modulated its expression. Furthermore, inhibition of miR-194 reversed the suppression of proliferation and invasion and the promotion of apoptosis induced by NEAT1 depletion in osteosarcoma cells. CONCLUSION: Knockdown of NEAT1 suppressed proliferation and invasion and induced apoptosis in osteosarcoma cells by inhibiting miR-194 expression.