Clinical Research of Detecting Plasma MiRNA-21 and MiRNA-143 for Identifying Early Esophageal Cancer and Benign Esophageal Diseases / 现代检验医学杂志

Objective To explore the clinical significance of detection of plasma microRNA-21,-143 in identifying early esophageal cancer and esophageal non-tumor diseases.Methods The expression of plasma microRNA-21,-143 in 27 cases of patients with early esophageal cancer (esophagus cancer group),25 cases of patients with non-esophageal tumor (non-esophageal tumor group)and in the healthy controls were detected by RT-PCR,and detected the levels of plasma CEA and CA72-4 by the electrochemical luminescence technology,which of changes were analysed to observe the relationship between the changes and the esophageal cancer,the benign esophageal diseases for the two markers.Results The expression of plasma microRNA-21,-143 in the esophagus cancer group were 0.93±0.17,0.27±0.05,which of ones in the non-esophagus cancer group were 0.25±0.03,0.99±0.15,and with those in the control group were 0.23±0.03,1.02±0.15.Compared with those in the non-esophagus cancer group,the expression of plasma microRNA-21,-143 were obviously up or down-regulated with significant differences (t=10.87,11.55,P＜0.01).Compared with those in the control group,which of ones were obviously up or down-regulated with significant differences (t=9.20,9.07,P＜0.01),and with no statistical significances in comparison between the esophagus cancer group and the control group (t=1.39,1.19,P＞0.05).The positiverate of plasma microRNA-21,-143 in the esophageal cancer,non-esophagus cancer group and the control group were,81.4 ％ (22/27),4.0 ％(1/25) and 0 (0/24);85.1％ (23/27),4.0％ (1/25),and 0 (0/24),respectively.The positive rate of microRNA-21,-143 in the esophageal group respectively in comparison with those in the non-esophagus cancer group and the control group were significantly higher,the differences had statistical significances (x2 =31.59,34.39,P＜ 0.01;x2 =34.42,37.23,P＜ 0.01).The expression of two markers in the esophagus cancer group were no statistically significant differences compared with control group (x2 =0.980,0.980,P＞0.05).The sensitivity and specificity of microRNA-21,-143 in early diagnosis on the esophageal cancer were 81.4 ％,97.9 ％ and 85.1％,97.9 ％.The sensitivity of microRNA-21,-143 in the esophageal group were significantly higher compared with those of CEA and CA72-4,the differences were statistically significant (x2 =12.79,P＜0.01;x2 =5.33,P＜0.05;x2 =15.03,P＜0.01;x2 =6.95,P＜0.05).The specificity of microRNA-21,-143 in the esophageal cancer group were no statistically significant differences in comparison with those of CEA and CA72-4 (x2 =1.043,0.000,P＞0.05) and (x2=1.043,0.000,P＞0.05),respectively.The analysis results from the spearman correlation test showed that in the esophageal cancer group,the expression of plasma microRNA-21,-143 had a negative correlation (r =0.658,P＜0.01).Which of ones respectively associated with the levels of CEA and CA72-4 (r=0.607,0.623,P＜0.01 and r=0.579,0.610,P＜0.01).Conclution The detection of expression of plasma miRNA-21,miRNA-143 in the patients with the early esophageal cancer and non-esophageal tumor can provide a new train of thought for pathologic diagnosis of early esophageal cancer.

MicroRNA-143 inhibits cell growth by targeting ERK5 and MAP3K7 in breast cancer

This study aimed to investigate the function and mechanism of microRNA-143 (miR-143) in the occurrence and development of breast cancer (BC). A total of 30 BC tissues, 30 corresponding noncancerous tissues, and 10 normal control (NC) breast tissues were obtained to detect the levels of miR-143, extracellular signal-regulated kinase 5 (ERK5) and mitogen-activated protein 3 kinase 7 (MAP3K7) using RT-qPCR, western blotting or immunohistochemistry. The correlation of miR-143 with ERK5 or MAP3K7 was evaluated using Pearson correlation analysis. MCF-7 cells were transiently transfected with miR-143 mimic, miR-143 inhibitor, miR-143 mimic/inhibitor + si-ERK5, si-MAP3K7 or si-cyclin D1. Then, cell growth was evaluated by MTT assay and the expressions of phospho-ERK5 (p-ERK5), ERK5, p-MAP3K7, MAP3K7 and cyclin D1 were detected by western blotting. Results showed that, compared with noncancerous tissues or NC breast tissues, miR-143 level was decreased, while p-ERK5, ERK5, p-MAP3K7 and MAP3K7 expressions were increased in BC tissues (all P<0.01). The miR-143 level was negatively correlated with the mRNA level of ERK5 or MAP3K7 (r=-4.231 or r=-4.280, P<0.01). In addition, up-regulated miR-143 significantly decreased the expressions of p-ERK5, ERK5, p-MAP3K7, MAP3K7 and cyclin D1 (all P<0.01), as well as cell viability in MCF-7 cells (all P<0.05) while the effect of down-regulated miR-143 was the opposite. In conclusion, both ERK5 and MAP3K7 may be the target genes of miR-143. Increased expression of miR-143 can inhibit cell growth, which may be associated with ERK5 and MAP3K7 expressions in BC.

Expression and Its Significance of MicroRNA-1 4 5 and MicroRNA-1 4 3 in Plasma in Children with Kawasaki Disease / 现代检验医学杂志

Objective To explore the diagnostic value of the quantitative detection of miRNA-145 and miRNA-143 in the ser-um of children with Kawasaki disease (KD).Methods In this study,45 KD cases were enrolled and were divided into 19 ca-ses with coronary artery lesions (CAL group)and 26 cases without coronary artery lesions (NCAL group).Thirty healthy children were recruited as the control group (NC group).qRT-PCR was conducted to detect the expression of miRNA-145 and miRNA-143 in serum of each group.The diagnostic value of miRNA-145 and miRNA-143 were evaluated by receiver op-erating characteristic curves (ROC)and the area under the curve (AUC)(95%CI).Results The relative expressions of miRNA-145 and miRNA-143 in the serum of the CAL group in acute phase were 2.33±1.26 and 1.64±0.50,which were respectively higher than the control group,with statistically significant difference (t=5.108,5.072,P0.05).During the acute phase of NCAL group, the relative expressions of miRNA-145 and miRNA-143 were 2.02±1.00 and 1.63±0.50 respectively.They were signifi-cantly higher than the control group (t=4.746,5.261,P0.05).The critical value of miRNA-145 for diagnosis of KD was 1.697 with sensitivity of 64.44% and specificity of 90% and yielded an area under the curve of ROC of 0.7733 (95%CI:0.667 0~0.879 7).Also,the critical val-ue of miRNA-143 was 1.361 with sensitivity of 64.44% and specificity of 86.67% and yielded an area under the curve of ROC of 0.8163 (95%CI:0.722 5~0.910 0)in discriminating KD from healthy group.Conclusion miRNA-145 and miR-NA-143 may prove to be a non-invasive biomarker for the auxiliary diagnosis of KD.

Effect of miRNA-143 on homocysteine induced vascular smooth muscle cell proliferation / 中国药理学通报

Aim To explore the effect of miRNA-143 ( miR-1 4 3 ) on homocysteine ( Hcy ) induced-vascular smooth muscle cells ( VSMCs ) proliferation and the mechanism .Methods VSMCs were cultured and in-cubated with Hcy by using primary cultured method . Then, cells were treated with different concentrations of Hcy and folate .VSMCs proliferation was determined with MTT assay , miR-143 was measured by qRT-PCR, and methylation of miR-143 was determined with meth-ylated PCR.Results After cells were treated with dif-ferent concentrations of Hcy , the proliferation of VSMCs was significantly increased , mRNA expression of miR-143 was decreased and methylation of miR-143 was increased .The proliferation of VSMCs was signifi-cantly decreased when transfected VSMCs with miR-143 precursor , and cell proliferation was increased by using miR-143 inhibitor transfection .Conclusion Hy-pomethylation of miR-143 may inhibit VSMCs prolifera-tion.

Inhibitory effect of miRNA-143 on the invasiveness of cervical can-cer cells by targeting MACC1 / 中国肿瘤临床

Objective:To illustrate the role of miRNA-143 on the invasiveness of cervical cancer cells. Methods:MiRNA-143 mimics or inhibitor sequences were transiently expressed in the cervical cancer cells by liposome transfection. Transwell assay was ap-plied to test the invasive ability of cervical cancer cells after miRNA-143 over-expression or inhibition. Bioinformatics assay was used to predict the targets of miRNA-143. RT-qPCR and luciferase reporter assay were performed to detect the expression of MACC1 mRNA in the cancer cells. RT-qPCR was conducted to test the expression of miRNA-143 and MACC1 mRNA in 20 fresh primary cervi-cal cancer and their matched para-neoplastic tissues. Statistical analyses were performed to evaluate the association between the expres-sion of miRNA-143 and MACC1 mRNA in the 20 cases of cervical cancer. Results:Transwell assays revealed that the miRNA-143 over-expression inhibited the cell invasiveness, while miRNA-143 inhibition promoted the invasive ability of the cervical cancer cells. Bioinformatics analyses revealed that miRNA-143 could target the 3'-UTR of MACC1. Dual luciferase reporter assay confirmed that miRNA-143 can affect 3'-UTR sequence in MACC1 genes. RT-qPCR analyses indicated that the expression of MACC1 mRNA was ob-viously down-regulated after miRNA-143 over-expression, while significantly increased after the miRNA-143 inhibition. The migration in Caski/miRNA-143 inhibitor cells was obviously elevated after being transfected with MACC1 shRNAs. RT-qPCR analyses showed that the expression of miRNA-143 was obviously decreased in the cancer tissues compared with the normal tissues, while MACC1 mRNA was apparently decreased in cancer tissues compared with the normal ones. Statistical analyses revealed that miRNA-143 was negatively correlated with MACC1 mRNA in the 20 cases of cervical cancer. Conclusion:This study reveals that miRNA-143 is down-regulated in the cervical cancer tissues. MiRNA-143 may play an important role in the regulation of cell invasiveness by targeting MACC1 in the cervical cancer cells.