RESUMO
OBJECTIVE: This study aimed to identify the different expression of microRNAs (miRNAs) in the plasma derived exosomes of patients with esophageal squamous cell carcinoma (ESCC). PATIENTS AND METHODS: A total of 9 patients with ESCC and 9 patients with benign diseases were involved. miRNA sequencing was performed to screen differential expression of microRNAs in plasma exosomes between patients with ESCC and controls. The function of miRNA on proliferation and migration abilities was determined by CCK-8 analysis, wound scratch and transwell test. Predicted target genes were screened by databases and confirmed by RT-qPCR. RESULTS: We identified a total of 10 miRNAs (7 upregulated and 3 downregulated) that were differentially expressed in plasma exosomes between patients with ESCC and control patients (fold change, FC ≥ 2.0 or ≤ -2.0, p ≤ 0.05) by miRNA sequencing. Ten miRNAs were detected by qRT-PCR to verify the results of the miRNA sequencing. MiR-103a-2-5p demonstrated the most significant differential expression in both exosomes of ESCC cell lines and plasma of patients as compared with control patients and was therefore selected for subsequent functional experiments. Overexpression of miR-103a-2-5p promoted proliferation and migration in TE-1 cells, whereas inhibition of miR-103a-2-5p suppressed proliferation and migration in KYSE-150 cells. Exosomes extracted from the cells transfected with miR-103a-2-5p mimics significantly increased the proliferation and migration of two ESCC cell lines. Two genes, CDH11 and NR3C1 were identified as predicted targets of miR-103a-2-5p by the bioinformatics tools TargetScan, MiRanda, and mirDIP and RT-qPCR. CONCLUSIONS: Our results shed light on how exosomal miR-103a-2-5p can promote proliferation and migration of ESCC cells and may represent a potential target for ESCC therapies.
Assuntos
Movimento Celular , Proliferação de Células , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/metabolismo , Carcinoma de Células Escamosas do Esôfago/patologia , MicroRNAs/metabolismo , Feminino , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-IdadeRESUMO
The hepatic stimulator substance (HSS) was partially purified from schedule-lighted weanling rat livers. The effects of HSS on p21(ras) expression of human hepatic carcinoma cell BEL-7402 were examined. The results showed: (1) HSS was capable of promoting the proliferation of the cells, and its molecular weight was 14 20 kD; (2) the p21(ras) expression was increased under HSS effect time-dependently, and the effect of HSS appeared synergistic with EGF; (3) the p21(ras) expression induced by HSS manifested a dose-dependent manner, and the saturation effect of HSS occurred at the concentration of 100 microgram/ml. Considering together with our previous report about HSS regulation on EGF receptor, these results strongly suggest that the proliferatory effect of HSS on hepatic carcinoma cells is presumedly related with EGF receptor-mediated signal transduction.
Assuntos
Carcinoma Hepatocelular/patologia , Substâncias de Crescimento/farmacologia , Neoplasias Hepáticas/patologia , Peptídeos/farmacologia , Proteínas Proto-Oncogênicas p21(ras)/biossíntese , Animais , Animais Recém-Nascidos , Carcinoma Hepatocelular/metabolismo , Divisão Celular/efeitos dos fármacos , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Neoplasias Hepáticas/metabolismo , Masculino , Mitógenos/farmacologia , Proteínas Proto-Oncogênicas p21(ras)/genética , Ratos , Ratos Sprague-Dawley , Células Tumorais CultivadasRESUMO
31 patients undergoing diagnostic cardiac catheterization for suspected congenital heart disease and 30 normal subjects were examined by pulsed Doppler echocardiography. Spectrum analysis of the shape of flow velocity curve and instantaneous flow velocities of the pulmonary artery during systolic period was made and their ratio calculated. The results showed that there was a remarkable difference between the flow velocity curve of pulmonary hypertension (PH) and that of normal pulmonary pressure. Combining with the characteristics of pulmonary arterial hemodynamics, a preliminary exploration was made on the mechanism of flow velocity curve changes. A ratio of flow velocities--PV% was proposed. The sensitivity and specificity for diagnosing PH with PV% were 94% and 100% respectively. There was a high correlation between PV% and pulmonary artery systolic pressure (r = -0.84) or pulmonary artery mean pressure (r = -0.82). It is concluded that PV% may eliminate many factors that influence the pulmonary artery hemodynamics and is a reliable, practical qualitative and quantitative index for diagnosing PH noninvasively.