ABSTRACT
Objective:To explore the effects of miRNA-373-3p (miR-373-3p) on the proliferation of nephroblastoma G401 cells through targeted regulation of CD44 expression.Methods:Bioinformatic method was used to predict the possible targeted genes of miR-373-3p based on bioinformatic databases including miRDB, miRanda, PITA and DIANA-microT. G401 cells were taken and transfected with miR-373-3p mimic, mimic negative control, miR-373-3p inhibitor or inhibitor negative control, respectively. Cell proliferation ability was detected by using CCK-8 assay. The number of clones was detected by using clone formation assay. The relative expression level of CD44 mRNA was detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression level of CD44 protein was detected by using Western blotting. The dual luciferase gene reporter assay was carried out in HEK-293T cells to vertify the target gene of miR-373-3p.Results:Bioinformatic analysis indicated that CD44 was a targeted gene of miR-373-3p. After 24 h transfection, the proliferation activity of G401 cells in miR-373-3p mimic group was decreased compared with that in mimic negative control group (all P < 0.05). After 48 h transfection, the proliferation activity of tumor cells in miR-373-3p inhibitor group was increased compared with that inhibitor negative control group (all P < 0.05). The formed number of clones in miR-373-3p mimic group was reduced compared with that in the mimic negative control group (55.3±2.5 vs. 90.7±2.9), and the difference was statistically significant ( t = 14.57, P < 0.01). The formed number of clones in miR-373-3p inhibitor group was more than that in inhibitor negative control group (115.0±2.7 vs. 92.0±2.4), and the difference was statistically significant ( t = 8.86, P < 0.01). The dual-luciferase gene reporter assay showed that CD44 was a direct targeted gene of miR-373-3p. The relative expression levels of CD44 mRNA in miR-373-3P mimic and mimic negative control group were 0.62±0.03 and 1.00±0.01, respectively, and the difference was statistically significant ( t = 11.28, P < 0.01). The relative expression levels of CD44 mRNA in miR-373-3p inhibitor and inhibitor negative control group were 1.31±0.02 and 1.00±0.00, respectively, and the difference was statistically significant ( t = 12.65, P < 0.01). The CD44 protein expression was decreased in miR-373-3p mimic group, while increased in miR-373-3p inhibitor group. Conclusion:miR-373-3p can inhibit tumor cell proliferation by targeting CD44 in nephroblastoma.
ABSTRACT
Objective To evaluate the effect of propofol (PPF) on stress response and lung injury in rats with traumatic brain injury (TBI).Methods A total of 36 SD rats were randomly (random number)divided into sham group,intralipid group,TBI group,PPF1 h group,PPF 2 h group,PPF 6 h group (n =6 in each group).Fluid percussion brain injury models were used.By intraperitoneal injection,intralipid was administered in intralipid group after sham operations,while propofol 100 mg · kg-1 was given to rats in PPF1 h group,PPF 2 h group and PPF 6 h group 1,2,6 hours following injury,respectively.Nerve motor function were scored at different intervals,serum concentrations of adrenocorticotropic hormone (ACTH),cortisol (COR) and norepinephrine (NE) were measured 12 h after injury.Seventy-two hours later,all rats were sacrificed and brains were harvested for TTC staining,and lungs taken were stained with HE staining for observation under light microscope and electron microscopy.Results Compared with sham and intralipid group,nerve motor function scores were significantly decreased,and serum concentrations of ACTH,COR and NE were increased significantly in rats after injury.Compared with TBI group,the above biomarkers were improved significantly after propofol injection.There were no significant differences in above biomarkersbetweenPPF 1 hand PPF 2 h group (t=-0.816,t=-0.208,t=0.582,P＞0.05).The differences in COR and NE concentrations between PPF 2 h and PPF 6 h group were statistically significant (t =3.018,P =0.013;t =3.662,P =0.004).Light microscopy demonstrated abundant inflammatory cell infiltration and massive thickening of the alveolar walls,Electron microscopy showed Type Ⅱ lung epithelial cell swelling,vacuolar degeneration,osmiophilic lamellar corpuscle emptying in cytoplasm,microvilli protrusions decreases in some cytoplasm in TBI group,and pathological damage was improved significantly in PPF 2 h group.Conclusions Propofol may inhibit stress and protect the lung tissue from damage in TBI rats.