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Objective:To investigate the expression of miR-211-5p in peripheral blood of patients with myelosuppression after neoadjuvant chemotherapy for breast cancer and its effect on Notch signaling pathway by targeting cyclooxygenase 2 (COX2) gene Regulation mechanism.Methods:From Jan. 2018 to Jan. 2021, 185 breast cancer patients who received neoadjuvant chemotherapy in Linyi People’s Hospital for the first time were included as the research objects. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the mRNA expression levels of miR-211-5p, COX2 gene and Notch signaling pathway related genes (Notch1, Jagged1 and Hes1) . The miR-211-5p mimic, inhibitor, mimic NC, and inhibitor NC were transfected into SD rat bone marrow mesenchymal stem cells, and the mRNA expression of miR-10a-3p and COX2 genes was detected by qRT-PCR for 48 hours. Western blot method was used to detect the protein expression levels of Notch signaling pathway related genes.Results:The relative expression of miR-211-5p in patients with severe myelosuppression was 2.41±0.32, which was significantly higher than that in patients with mild myelosuppression (1.53±0.18) ( t=6.385, P<0.001) ; The relative expression of COX2 gene mRNA in patients with severe myelosuppression was 3.64±0.74, which was significantly lower than that in patients with mild myelosuppression (5.37±1.02) ( t=7.469, P<0.001) . In patients with severe myelosuppression, there was a significant negative correlation between miR-211-5p and COX2 gene mRNA levels ( r=-0.694, P=0.006) . The results of the dual luciferase report experiment confirmed that COX2 was the target gene of miR-211-5p. The relative expression of Notch1, Jagged1, and Hes1 mRNA in peripheral blood of patients with severe myelosuppression were 2.35±0.41, 2.76±0.46 and 3.04±0.52, respectively, which were significantly lower than those in patients with mild myelosuppression (4.12±0.63, 4.53±0.58 and 5.12±0.67) ( t=5.367, 6.114 and 6.135, respectively, P<0.001) . After transfecting SD rat bone marrow mesenchymal stem cells with miR-211-5p mimics and inhibitors, the relative expression of miR-211-5p in the mimic group was 3.46±0.49, significantly higher than that in the mimic NC group (2.24±0.32) The relative expression of miR-211-5p in the inhibitor group was (1.28±0.19) and (2.33±0.37) inhibitor NC group ( P<0.001) , while the relative expression of miR-211-5p in the inhibitor group was significantly lower than that in the other three groups ( P<0.001) . The mRNA expression of COX2 gene in mimic group was 2.73±0.36, which was significantly lower than that in mimic NC group (4.05±0.59) , inhibitor group (6.15±0.86) and inhibitor NC group (4.18±0.65) ( P<0.001) , while mRNA expression of COX2 gene in the inhibitor group was significantly higher than that in the other three groups ( P<0.001) . The expression of Notch1, Jagged1, and Hes1 in the inhibitor group was significantly increased, while the expression of Notch1, Jagged1, and Hes1 in the mimic group was significantly decreased. Conclusion:The expression level of miR-211-5p in peripheral blood of severe myelosuppressed patients with breast cancer after neoadjuvant chemotherapy is significantly increased, and the Notch signaling pathway can be inhibited by targeted down-regulation of COX2 gene expression.
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OBJECTIVE@#To investigate the expression and correlation of miR-211, miR-155, and C-myc in acute T lymphocytic leukemia (T-ALL), aiming to provide evidence for the diagnosis and treatment.@*METHODS@#A total of 96 T-ALL patients who were diagnosed and treated in People's Hospital of Zhengzhou from June 2014 to June 2017 were selected, and 69 healthy volunteers who had a physical examination were selected as control group in the same period. Real-time fluorescent quantitative PCR (RT-PCR) was used to determine the expression levels of miR-211, miR-155, and C-myc in peripheral blood mononuclear cells in each group. Kaplan-Meier was used to analyze the survival of T-ALL patients and correlation of miR-211, miR-155, and C-myc with prognosis. Pearson correlation analysis was used to evaluate the correlation of miR-211, miR-155, and C-myc with disease risk.@*RESULTS@#The expression levels of miR-211 mRNA, miR-155 mRNA, and C-myc mRNA in T-ALL group were higher than those in the control group (P<0.01), those in non-remission group were higher than those in remission group (P<0.01), and those in high-risk group were also higher than those in low-risk group and intermediate-risk group (P<0.01). The survival time of T-ALL patients with low miR-211 expression was longer than that with high miR-211 expression (P<0.01), that with low miR-155 expression was longer than that with high miR-155 expression (P<0.01), and that with low C-myc expression was also longer than that with high C-myc expression (P<0.01). The high expression of miR-211, miR-155, and C-myc was linearly positively correlated with high risk of disease (r=0.749, 0.781, 0.804).@*CONCLUSION@#The expressions of miR-211, miR-155, and C-myc are up-regulated in T-ALL patients, closely related to prognosis, and linearly positively correlated with disease risk.
Subject(s)
Humans , Leukocytes, Mononuclear , MicroRNAs/genetics , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/genetics , Prognosis , Proto-Oncogene Proteins c-myc/genetics , RNA, MessengerABSTRACT
Objective:To investigate the relationship between miR-211 and the occurrence of epithelial ovarian cancer,and its influence on the proliferation of ovarian cancer cells. Methods:To analyze the expression miR-211,CDK6 and Cyclin D1 of 30 cases of ovarian cancer and ovarian cancer cell lines,and 30 cases of non-ovarian cancer tissues and the normal ovarian epithelial cells were selected as the control group,and to analyze effects of miR-211 on the proliferation of ovarian cancer cells,as well as the Cyclin D1 and CDK6. Results:miR-211 relative expression level of ovarian cancer tissue was significantly lower than that in the normal group ( P<0. 05). Relative expression level of miR-211 of ovarian cancer cells was significantly lower than that in normal ovarian epithelial cells (P< 0. 05);in epithelial ovarian cancer cell line HO8910,cell number of miR-211 on the 3-day and the 4-day was significantly lower than that of miR-Ctrl group (P<0. 05);relative expression levels of Cyclin D1 and CDK6 in epithelial ovarian cancer were significantly higher than those in normal ovarian epithelial tissues (P<0. 05);miR-211 of epithelial ovarian cancer cell lines significantly inhibited Cyclin D1 and CDK6 expression;in ovarian cancer tissues,Spearman correlation analysis results showed that relative expression levels of miR-211 and Cyclin D1 and CDK6 was negatively correlated ( r=-0. 583, P= 0. 010 ) . Conclusion: miR-211 can inhibit the proliferation of ovarian cancer cells,and inhibit the expression of Cyclin D1 and CDK6;miR-211,Cyclin D1 and CDK6 in ovarian cancer may be involved in the regulation of ovarian cancer.
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Objective To investigate the role of miR-211/TFAM in the regulation of proliferation of breast cancer cells .Methods In the present study , we transfected breast cancer cells with miR-211 mimics or miR-211 inhibitor to achieve miR-211 and detected the expression of miR-211 and the expression level of TFAM proteins in response to miR-211 overexpression or miR-211 silencing;luciferase reporter gene plasmid with or without a six base pairs mutation in the 3′UTR of TFAM ( mut-TFAM/wt-TFAM) were conducted and co-transfected with miR-211 mimics or miR-211 inhibitor, then the change of the luciferase activity was detected;then pcDNA3.1/TFAM plasmid was constructed and co-transfected with miR-211 mimics or miR-211 inhibitor, TFAM protein expression level changes were determined in response to miR-211 overexpression or miR-211 silencing; lastly the cell proliferation was determined in response to mimics NC/miR-211 mimics and pcDNA3.1/TFAM co-transfection.Results Overex-pression of miR-211 inhibits the expression of TFAM protein , miR-211 silencing promote TFAM protein expression;miR-211 can regulate the expression of TFAM by direct targeting;TFAM over expression was achieved by pcDNA3.1/TFAM transfection , and TFAM overe xpression can restore the inhibitory effect of miR-211 on TFAM;miR-211 inhibited the growth and proliferation of breast cancer cells , TFAM can promote the growth and proliferation of breast cancer cells;TFAM restored the inhibitory effect of miR-211 on growth and proliferation of breast cancer cells .Conclusions miR-211 regulates the growth of breast cancer cell with targeting of HMGB .
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Objective To study the effect of Sirtuin1 (Sirt1) on the pathological process through its activity of deacetylation to improve the clinical outcome of acute sepsis. After searching data base, microRNA-211 (miR-211) was found to have action potentially targeting at Sirt1.The present study aimed to find the interaction between miR-211 and Sirt1 in the pathogenesis of hypoxic injury to cardiomyocytes in the presence of lipopolysaccharide ( LPS ) . Methods Primary neonatal rat cardiomyocytes ( NRC ) isolated from neonatal SD rats and H9c2 ( cardiomyocytes after culture with 10% fetal serum of cattle and DMEM under 37 ℃ and 5% CO2 ) cell line were used in the experiments.The miR-211 expression was quantified by qRT-PCR after LPS exposure for 4 hours, and the changes in Sirt1 protein level were also detected in both NRC and H9c2 by western blot.At the same time, CCK-8 assay and TUNEL staining were also performed to measure cell proliferation and apoptosis activation when either treated with LPS alone or followed by exposure to hypoxia.Results Compared to the control group, the doses of 20μg/mL, 40μg/mL LPS treatment for 4 hours had no significant effects on H9c2 cell proliferation at 24 h, 48 h, 72 h, but it could significantly induce the cell apoptosis of neonatal rat cardiomyocytes and H9c2 cells after hypoxia, and the apoptosis rate increased all over 100% in both NRC and H9c2.At the same time, LPS treatment could significant up-regulate miR-211 expression which was closely associated with decrease in Sirt1 protein levels.Conclusions LPS enhanced cardiomyocytes injury after exposure to hypoxia which was closely associated with up-regulating miR-211 expression and in turn to suppress Sirt1 expression.