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OBJECTIVE@#To analyze the association of integrinα5 (ITGA5) with grading of liver cancer and the overall patient survival and investigate the effects of integrin α5 (ITGA5) silencing on the proliferation, invasion and migration abilities of human liver cancer Bel-7404 cells.@*METHODS@#UALCAN was used to analyze the expression of ITGA5 in liver cancer tissues and normal tissues, and expression in different grades of liver cancer tissues. GEPIA was used to analyze the relationship between ITGA5 expression and the survival of liver cancer patients through.The ITGA5 shRNA lentiviral vector was used to infect Bel-7404 cells to establish a cell line with stable ITGA5 silencing verified by Western blotting. Plate clone formation assay and Transwell assay were used to detect the proliferation, invasion and migration of Bel-7404 cells. The correlation between ITGA5 and PI3K in liver cancer tissues and control tissues was analyzed using Oncomine cancer specimen database.@*RESULTS@#The expression of ITGA5 was significantly higher in liver cancer than in normal tissues ( < 0.05). The expression of ITGA5 was significantly lower in grade 1 than in grade 2 liver cancer, and also lower in grade 2 than in grade 3 liver cancer ( < 0.05). The patients with high ITGA5 expression had a significantly lower overall survival rate than those with low ITGA5 expression ( < 0.05). Plate clone formation assay showed that the clone formation rate was significantly lowered in Bel-7404 cells with ITGA5 silencing compared with the blank and negative control cells ( < 0.05). ITGA5 silencing significantly attenuated the migration of Bel-7404 cells as shown by Transwell assay ( < 0.05). ITGA5 and PI3K were both highly expressed and showed a positive correlation in liver cancer tissues ( < 0.05).@*CONCLUSIONS@#ITGA5 is closely related to the progression of liver cancer and the patients' prognosis. ITGA5 silencing inhibits the proliferation, invasion and migration of liver cancer cells. ITGA5 promotes the liver cancer growth and metastasis possibly by regulating the PI3K signaling pathway.
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Humans , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Integrin alpha5 , Liver Neoplasms , Neoplasm Invasiveness , Phosphatidylinositol 3-KinasesABSTRACT
Objective To investigate the changes and significance of serum anti-CCP (ACCP)antibody,matrix metalloprotein-ase-3 (MMP-3 ),interleukin-17 (IL-17 ),interleukin-18 (IL-18 )in the patients with rheumatoid arthritis (RA).Methods The ELISA method was adopted to detect the in the peripheral blood serum ACCP antibody,MMP-3,IL-17 and IL-18 in 80 patients with RA,32 patients with osteoarthritis (OA)and 32 cases of healthy controls and the detection results were performed the statis-tical analysis.Results The ACCP antibody,MMP-3,IL-17 and IL-18 levels in the RA group were significantly higher than those in the OA group and the healthy control groups;the ACCP antibody,MMP-3,IL-17 and IL-18 levels in the low,middle and high RA activity groups were higher than those in the stable group,the ACCP antibody,MMP-3,IL-17,IL-18,disease activity score (DAS28),C-reactive protein (CRP)and erythrocyte sedimentation rate(ESR)in the RA group were increased;the MMP-3,IL-17, IL-18 and DAS28 in the ACCP antibody positive group were higher than those in the A CCP antibody negative group;the positive correlation existed among the ACCP antibody,MMP-3,IL-17 and IL-18 in the RA group (P <0.05);the ACCP antibody,MMP-3, IL-17 and IL-18 were positively correlated with the monitoring indicators of CRP and DAS28 in the low,middle and high RA activi-ty groups.Conclusion MMP-3,IL-17 and IL-18 participate in the occurrence and development process of RA;The detection of ser-um ACCP antibody,MMP-3,IL-17 and IL-18 has a certain value in the judgment of disease activity,and prevention and treatment in the patients with RA.
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Objective:In order to investigate the role of microRNA in the pathogenesis of ankylosing spondylitis patients,we detected the peripheral blood of patients with ankylosing spondylitis in miR-17,miR-181,miR-106,miR-30 and miR-495 expression, thereby to study the role of microRNA regulation of clinical and diagnostic value in the pathogenesis of ankylosing spondylitis provide new ideas.Methods:Collection of patients with ankylosing spondylitis and normal peripheral blood,peripheral blood mononuclear cells were isolated and extracted PBMC small RNA,using primers specific stem-loop reverse transcribed into cDNA,and build a mature miR-17,miR-181,miR-106,miR-30 and miR-495 T-carrier,standard curve,the use of stem-loop method by Real-time PCR technology to detect miR-17,miR-181,miR-106,miR-30 and miR-495 expression level.Results: In this study,92 cases of clinical samples were collected,of which 61 patients with AS,normal 31 cases.By Real-time PCR detection showed that compared with normal subjects,there was upregulation of miR-106 (P>0.05) and miR-30 (P>0.05) in the peripheral blood of patients with ankylosing spondylitis;down-regulation were miR-181 ( P0.05 ) , in which the miR-181 and miR-495 was statistically significant.Conclusion:Compared with normal, there are differences in the peripheral blood of patients with ankylosing spondylitis expression of miR-495 and miR-181,which may be targeted to regulate TLR-4,HLA-B,DVL,GSK/3βgenes,this may be found in the pathogenesis of ankylosing spondylitis studies provide new ideas,to become a new clinical diagnosis markers.
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Objective:To investigate the expression of miR-16,miR-17,miR-30a etc in peripheral blood plasma ,mononuclear cells ( PBMC) and synovial fluid of patients with rheumatoid arthritis ( RA) and its clinical significance ,to provide a theoretical basis for the further research in the mechanism of RA.Methods:Collected 80 cases of RA patients ,32 cases of osteoarthritis ( OA) patients and 32 healthy human peripheral blood ,synovial fluid ,separating the plasma and PBMC;extraction of small RNA ,with specific stem loop primed reverse transcription into cDNA , establishing a mature miRNA-T carrier and making standard curve;stem-loop method Real-time quantitative PCR was adopted to detect the expression of miRNAs in plasma ,PBMC and synovial fluid,and for correlation analysis;and with RA activity monitoring indicators rheumatoid factor (RF),erythrocyte sedimentation rate (ESR),C-reactive protein (CRP) correlation analysis.Results:In RA group,the expression of miR-16,miR-17,miR-30a,miR-106,miR-101 and miR-130 in plasma,PBMC and synovial fluid were upregμlation compared with OA group,the healthy control group (P<0.05),but the expression level of miR-101 in plasma of RA and OA -group difference was not statistically significant.Correlation analysis showed that 6 kinds of miRNA in the plasma ,PBMC and joint fluid have varying degrees of positive correlation ( P<0.05 ).Correlation analysis showed that miRNA in plasma , PBMC and synovial fluid have one or more indicators of a positive correlation with RF , ESR, CRP ( P<0.05 ).Conclusion:The expression of miR-16,miR-17,miR-30a,miR-101,miR-106 and miR-130 is changes significant in the peripheral blood and synovial fluid of RA patients ,indicate the miRNAs may be through some related genes play an important role in the process of the pathogenesis of RA;its expression level can be used as an effective indicator of RA disease activity , and provide new diagnostic markers for RA.
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To detect the influence of rapamycin on the expression of miR-30b,miR-200a and miR-17-5p etc in macrophages and provide the basis to study the regulation of miRNA in autophagy mechanism of macrophages .Methods: Small RNA was extracted at different times after stimulated with rapamycin in cultured RAW 264.7 cells.After using the stem-loop reverse transcription primers to reverse transcribed into cDNA ,the expression of miR-30b ,miR-30c,miR-106a,miR-214,miR-183,miR-200a, miR-376c,miR-17-5p, miR-142-3p, miR-377 was detected by Real-Time PCR.Results: After RAW264.7 cells was treated by rapamycin for 2,4,6 and 8 hours,the expression of miR-17-5p and miR-106 increased (More than 2.1 times,P<0.05) in 2,4 and 6 hours;miR-214 was up regulated in 2 and 8 hours (More than 2.4 times,P<0.05);miR-30b,miR-30c,miR-183,miR-200a,miR-376c and miR-142-3p was up regulated in 2,6 and 8 hours (2.4 times,P<0.05 );while miR-183 and miR-200a was down regulated at 4 hours(More than 2.1 times,P<0.05);miR-30b was significantly low expression in 8 hours (more than 50 times,P<0.05);miR-377 was up regulated at 4 hours (more than 2.5 times,P<0.05),but was significantly down regulated at 2 and 8 hours (More than 50 times,P<0.05) Conclusion: The expression of miR-200a,miR-30b,miR-377,miR-30c,miR-376c and miR-17-5p is significantly changed after rapamycin stimulated RAW264.7 macrophages,indicated the miRNA may plays an important role in autophagy through the regulation of autophagy-related genes.