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Parkinson's disease(PD)is the second most common neurodegenerative disease after Alzheimer's disease,with high morbidity and high disability rate.Since the early symptoms of PD are not typical and often similar to those of normal aging or other diseases.It is easy to missed diagnosis and misdiagnosis,which seriously affects the diagnosis and treatment of this disease and aggravetes the burden on the patients' life.MicroRNAs(miRNA)are a class of endogenous non-coding RNAs that are involved in post-transcriptional regulation by binding to target messenger RNAs(mRNA).They are highly conserved,short,easy to obtain,and can stably exist in peripheral body fluids.They have been used as biomarkers for a variety of diseases.Recent studies have demonstrated that miRNA play an important role in the development of PD.This paper reviews the recent research progress of miR-7/124/155,three mature miRNA in PD,aiming to provide reference for clarifying the pathogenesis and guiding the diagnosis and treatment of PD.
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Humans , Parkinson Disease , Neurodegenerative Diseases , MicroRNAs/genetics , Gene Expression Regulation , Biomarkers/metabolismABSTRACT
Objective To detect the expression levels of miR-7-5p and POLE4 in non-small cell lung cancer cells and their effect on cells proliferation, migration and invasion. Methods qRT-PCR was used to detect the relative expression levels of miR-7-5p and POLE4 mRNA in NSCLC tissues, adjacent tissues, tumor cells and human normal bronchial epithelial cells. Luciferase reporter gene was used for analyzing of the targeting relation between POLE4 and miR-7-5p in NSCLC cells. si-NC and si-POLE4 were transfected into SPC-A-1 cells as the si-NC group and si-POLE4 group, and the control group was set at the same time. MTT method, scratch test and Transwell test were used to detect cell proliferation, migration and invasion. Results The expression levels of miR-7-5p in NSCLC tissues and cells were reduced, and the expression levels of POLE4 were increased. miR-7-5p could target to combine with POLE4. After 72 hours of culture, the OD value in si-POLE4 group was significantly lower than those in the control group and si-NC group (P < 0.05). The migration rate and the number of transmembrane cells in the si-POLE4 group cultured for 48 hours were lower than those in the control group and the si-NC group (P < 0.05). Conclusion miR-7-5p may inhibit the proliferation, migration and invasion of non-small cell lung cancer cells by targeting POLE4.
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Objective MicroRNAs are differentially expressed in colorectal cancer tumor tissues and adjacent normal tissues, which play an important role in the development of colorectal cancer. This study aims to investigate the expression of microRNA-7-5p in colorectal cancer patients and its influence on the proliferation and apoptosis of colon cancer cell CaCo2. Methods The high-throughput microarray was used to screen differentially expressed microRNAs, and real-time quantitative PCR (RT-PCR) was used to detect the expression of microRNA-7-5p in 10 cases of colorectal cancer patients and corresponding adjacent normal tissues. Western blot was performed to detect the expression of intestinal trefoil factor (TFF3) in different tissues and CaCo2 cells after transfection with microRNA-7-5p. The expression of TFF3 in different tissues and CaCo2 cells transfected with microRNA-7-5p was detected. TUNEL combined with flow cytometry was used to detect the apoptosis of CaCo2 cells after transfection. The CCK-8 assay was used to detect the proliferation of CaCo2 cells after transfection. Results The relative expression of MicroRNA-7-5p in colorectal cancer tissue was 0.409 ± 0.095, which was significantly lower than that of normal tissue adjacent to cancer (1.000 ± 0.014), and the difference was statistically significant (P <0.01). The relative expression of TFF3 in colorectal cancer tissues was significantly higher than that in normal adjacent tissues (P <0.01). The relative expression of TFF3 in CaCo2 cells decreased in the overexpressed microRNA-7-5p of the control group (0.729 ± 0.041). The proliferation ability (0.930 ± 0.007) was significantly lower in the blank control group (0.990 ± 0.005) (P <0.01), and it could increase the proportion of early apoptotic cells (50.700 ± 0.989) and late apoptotic cells (40.525 ± 0.515). Conclusion MicroRNA-7-5p is lowly expressed in colorectal cancer and is associated with the occurrence and development of colorectal cancer. Up-regulated microRNA-7-5p inhibits the proliferation of colon cancer cell CaCo2 and promotes its apoptosis.
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Objective@#To investigate the effects of overexpression of microRNA-7 (miR-7) on the proliferation and invasion of HepG2 cells and the underlying mechanism in vitro.@*Methods@#The relative expression levels of miR-7 and Raf1 in hepatocellular carcinoma (HCC) tissues and adjacent normal tissues (ANT) were detected by quantitative real time-PCR (qRT-PCR). The relationship between the expression of miR-7 and the characteristics of HCC patients was analyzed. Cells were divided into blank control group, negative control (NC) group and miR-7 mimics transfected group, miR-7 mimics and NC were transfected into HepG2 cells by Lipofectamine™2000. The relative expression of miR-7 was detected by qRT-PCR. The proliferation ability of HepG2 cells was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The invasion of HepG2 cells was detected by Transwell assay. The target genes of miR-7 were predicted by TargetScan and the binding effect of miR-7 on the 3′UTR of Raf1 was verified by dual luciferase reporter assay.The expressions of Raf1 protein in hepatocellular carcinoma tissues, normal tissues and miR-7 mimics transfected HepG2 cells was detected by Western blot. The correlation of the levels of miR-7 and Raf1 mRNA was determined by Pearson correlation analysis.@*Results@#The relative expression level of miR-7 in HCC was 0.49±0.02, significantly lower than in ANT (1.21±0.05, P<0.01). The level of miR-7 was significantly correlated the tumor volume, metastasis and prognosis of HCC patients (P<0.05). The relative expression level of miR-7 in miR-7 mimics transfected HepG2 group was 12.67±0.40, significantly higher than that in blank group (P<0.01). Compared with the blank group, the A value and invasion ability of miR-7 mimics transfected group were significantly down-regulated at 48 hours and 72 hours after transfection (P<0.01). Compared with miR-7 NC group, the luciferase activity of wild-type Raf1 reporter gene in miR-7 mimics transfected group was significantly reduced (P<0.01). The relative expression of Raf1 protein in HCC was 3.15±0.10, significant higher than in ANT (0.53±0.03, P<0.01). The relative expression of Raf1 protein in miR-7 mimics transfected group was 0.24±0.01, significantly lower than in miR-7 NC group (0.98±0.02, P<0.01). Furthermore, an negative correlation was observed between the levels of miR-7 and Raf1 in HCC tissues (P<0.05).@*Conclusions@#The expression of miR-7 in HCC is significantly decreased and inversely correlated with poor survival of HCC patients. Overexpression of miR-7 can inhibit the proliferation and invasion ability of hepatocellular carcinoma cells HepG2 by downregulating Raf1 in vitro.
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AIM:To study the effect of microRNA-7 (miR-7) knockdown (KD) on concanavalin A (ConA)-induced acute liver injury (ALI) in mice.METHODS:Wild type (WT) mice and miR-7KD mice were received ConA (30 mg/kg) to induced acute liver injury model by intraperitoneal injection,and the morphological changes,liver weight and weight index were measured 48 h later.The pathological changes of the liver tissues were observed by HE staining.The levels of serum alanine aminotransferase (ALT),IL-4 and IFN-γ were detected by ELISA.The proportional changes of CD4 + T cells and the relative levels of IL-4 and IFN-γwere analyzed by flow cytometry.RESULTS:The color of the liver tissue became lighter,and the weight and weight index were changed significantly in miR-7KD mice compared with control group (P ＜ 0.05).HE staining showed that the inflammatory cell infiltration was increased in the liver of miR-7KD mice.Moreover,the level of serum ALT was significantly increased (P ＜ 0.05).The serum level of IFN-γelevated significantly (P ＜ 0.01),while the IL-4 levels decreased significantly (P ＜ 0.01) in the serum of miR-7KD mice.Furthermore,the proportion of CD4 + T cells and relative IFN-γcells increased obviously (P ＜ 0.01).CONCLUSION:miR-7 knockdown promotes the pathogenesis of the ConA-induced acute liver injury in mice.
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AIM:To study the effect of microRNA-7 (miR-7) knockdown (KD) on concanavalin A (ConA)-induced acute liver injury (ALI) in mice.METHODS:Wild type (WT) mice and miR-7KD mice were received ConA (30 mg/kg) to induced acute liver injury model by intraperitoneal injection,and the morphological changes,liver weight and weight index were measured 48 h later.The pathological changes of the liver tissues were observed by HE staining.The levels of serum alanine aminotransferase (ALT),IL-4 and IFN-γ were detected by ELISA.The proportional changes of CD4 + T cells and the relative levels of IL-4 and IFN-γwere analyzed by flow cytometry.RESULTS:The color of the liver tissue became lighter,and the weight and weight index were changed significantly in miR-7KD mice compared with control group (P ＜ 0.05).HE staining showed that the inflammatory cell infiltration was increased in the liver of miR-7KD mice.Moreover,the level of serum ALT was significantly increased (P ＜ 0.05).The serum level of IFN-γelevated significantly (P ＜ 0.01),while the IL-4 levels decreased significantly (P ＜ 0.01) in the serum of miR-7KD mice.Furthermore,the proportion of CD4 + T cells and relative IFN-γcells increased obviously (P ＜ 0.01).CONCLUSION:miR-7 knockdown promotes the pathogenesis of the ConA-induced acute liver injury in mice.
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Objective To investigate the expression and clinical significance of microRNA‐7 (miRNA‐7) and focal adhesion kinase (FAK) in colorectal cancer (CRC) .Methods Sixty pairs of CRC and adjacent colorectal tissues were collected .The expression of FAK was detected by immunohistochemistry and the expression of miRNA‐7 was determined by quantitative real‐time polymerase chain reaction (PCR) . Chi square test was used for statistical analysis and Spearman rank was applied for correlation analysis . Results The positive rate of FAK expression in CRC was 75 .0% (45/60) and that in adjacent colorectal tissues was 26 .7% (16/60) ,the difference was statistically significant (χ2 = 28 .04 , P < 0 .01) . The positive rate of phospho‐FAK (p‐FAK ) expression in CRC was 65 .0% (39/60 ) and that in adjacent colorectal tissues was 21 .7% (13/60) ,and the difference was statistically significant (χ2 = 22 .94 , P <0 .01) .The expression of miRNA‐7 in CRC tissues was down‐regulated compared with that in adjacent colorectal tissues (P= 0 .044) .The correlation between miRNA‐7 expression and lymph nodes metastasis was negative in patients with CRC (Z= - 2 .290 ,P= 0 .022) .The miRNA‐7 expression was significantly negatively correlated with TNM stage in patients with CRC (Z= - 2 .698 , P= 0 .007) .However it was not correlated with age ,gender ,the location of tumor and tumor differentiation .The correlation between miRNA‐7 expression and FAK ,p‐FAK was negative (Z= - 0 .303 ,P= 0 .019 ;Z= - 0 .267 ,P= 0 .038) . Conclusion The miRNA‐7 may involved in the genesis and development of CRC through regulating the expression of FAK ,which provide a new target for the diagnosis and treatment of CRC .
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Objective To detect the expressions of microRNA-126 (miR-126) and microRNA-7 (miR-7) in esophageal squamous cell carcinoma (ESCC) and to analyze their correlations with clinicopathologic features and prognosis of ESCC.Methods The expressions of miR-126 and miR-7 in 116 ESCC samples and matched normal tissue samples were detected by real-time PCR.Statistical analysis was used to find the relationships among the expressions of miR-126 and miR-7,pathological characteristics and prognosis.Results Low expression,normal expression and high expression of miR-126 were found in 73 (62.9％),35 (30.2％) and 8 (6.9％) carcinoma samples respectively.Low expression,normal expression and high expression of miR-7 were found in 52 (44.8％),35 (30.2％) and 29 (25.0％) carcinoma samples respectively.The disease-free survival in patients with low expression of miR-126 and miR-7 was shorter than that in patients with non-low expression (x2 =4.268,P ＜0.05 ; x2 =4.993,P ＜0.05).The low expression of miR-126 was correlated with tumor location,family history and drinking (x2 =14.564,P ＜ 0.05 ; x2 =5.691,P ＜ 0.05 ; x2 =4.971,P ＜ 0.05),but was uncorrelated with gender,age,diferentiation,infiltration,lymphatic metastasis and smoking (all P ＞ 0.05).The low expression of miR-7 was uncorrelated with pathological characteristics of ESCC (all P ＞ 0.05).Conclusion The low expressions of miR-126 and miR-7 may be related to the prognosis of patients with ESCC,and have a certain clinical detection significance.
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Objective To construct a lentiviral vector containing mir-7-3 gene and green fluorescent protein (GFP) gene,and to detect the expression of mir-7-3 gene in U251 cells.Methods The fragments containing all the mir-7-3 gene were amplified by RT-PCR and were cloned into the lentivirus vectors labeled with GFP,which was transfected together with the packaging plasmids into 293T cells by CaC12.The supernatant was collected,concentrated,identified,and was transfected to U251 cells of gliomas.Fluorescent microscopy was used to observe the fluorescence in the 293T cell,and real time RT-PCR was used to examine the relative contents of mir-7-3 in U251 cells.Results Electrophores was shown that the sequence of the RT-PCR product was consistent with the data of mir-7-3 by DNA sequence analysis,indicating that the mir-7-3 gene was successfully cloned,and strong green fluorescence was observed by fluorescent microscopy.The supernatant of lentivirus-transfected 293T cells effectively infected U251 cells and the relative content of mir-7-3 was observed in the transfected U251 cells.Conclusion It is concluded that the lentiviral vector containing mir-7-3 gene was constructed successfully,which provides a basis for further study of mir-7-3 function.