ABSTRACT
ObjectiveTo investigate the protective effect of microRNA-155 (miR-155) antisense oligonucleotid (ASO) on acute lung injury (ALI) mice by establishing a lentiviral expression vector of ASO of miRNA.Methods miR-155 antisense oligonucleotides amplified by polymerase chain reaction (PCR) from genomic, using BamH Ⅰ and Nhe Ⅰ double digestion, ligated into lentiviral expression vector. Sequence and virus titer were measured. According to the random number table method, 54 male BALB/c mice of 4-6 weeks old were divided into three groups. ALI animal models were prepared by intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS). The three groups were injected with 200μL phosphate buffered saline (PBS) containing 1×108/mL pmiR-155-ASO virus (pmiR-155-ASO group) or 200μL PBS containing 1×108/mL pSMPUW-miR-GFP empty virus (pmiR-cont group) or the same amount of PBS (PBS group) at 24 hours before the molding. Ten mice in each group were used to observe the 7-day survival rate. Blood samples and lung tissues of the remaining 8 mice were harvested after the model was established, and the levels of serum inflammatory cytokines were determined by enzyme linked immunosorbent assay (ELISA); the expression of miR-155 in lung tissue was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR); histopathological changes of lung and distribution of macrophages were observed under microscope.Results There was no significant difference in each index between pmiR-cont group and PBS group. The mature miR-155 expression in lung tissue in pmiR-155-ASO group was significantly lower than that in pmiR-cont group (2-ΔΔCt: 4.92±0.72 vs. 15.38±0.60,P < 0.05). Compared with pmiR-cont group, the injury degree of ALI mice after pretreatment with miR-155ASO was significantly improved, and the 7-day survival rate was significantly increased (72.1％ vs. 61.9％,P < 0.05 ); gross lung observation showed that congestion in lung tissue was significantly reduced, and the ratio of wet/dry weight (W/D) of lung was significantly decreased (4.50±0.13 vs. 5.64±0.61,P < 0.05);hematoxylin-eosin (HE) staining showed that inflammatory cell infiltration in lung tissue was decreased, while immunofluorescence assay showed that macrophage infiltration in lung tissue was significant decreased; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL- 6) in serum were significantly decreased [TNF-α (ng/L):379.8±48.9 vs. 495.9±33.3, IL-6 (ng/L): 262.3±61.8 vs. 355.4±22.6, bothP < 0.05], but the level of IL-10 did not change significantly (ng/L: 143.6±32.5 vs. 140.4±22.3,P > 0.05).Conclusion miR-155 ASO has the effect of inhibiting LPS-induced inflammatory response and improving prognosis in ALI mice.
ABSTRACT
Objective To investigate the role of combined analysis of E2F3a and CASP8AP2 expression in prognosis evaluation in pediatric acute lymphoblastic leukemia (ALL). Methods The study included 141 newly diag-nosed pediatric ALL patients enrolled at the Hematology Center,Beijing Children′s Hospital,Capital Medical Universi-ty between March 2008 and July 2010,including 97 boys and 44 girls(aged 1. 2 - 15. 5 years,median 5. 2 years). E2F3a and CASP8AP2 expressions were quantified in 141 children with ALL by adopting real - time quantitative poly-merase chain reaction (qPCR). Receiver operating characteristic (ROC)curve was used to find the best cut - off point to divide the patients into E2F3a or CASP8AP2 low - and high - expression groups,and the treatment outcome between the groups was compared. Cox regression was used to analyze the prognostic significance of the combined expression of E2F3a and CASP8AP2. Results The estimated 5 - year relapse free survival(RFS)rate,event free survival(EFS) rate and overall survival (OS)rate of patients with low - E2F3a and low - CASP8AP2 expression were (58. 9 ± 10. 0)％,(56. 0 ± 9. 9)％ and (72. 0 ± 9. 0)％,respectively. They were significantly lower than those of patients with high - E2F3a and high - CASP8AP2 expression,whose RFS,EFS and OS were (94. 9 ± 2. 5)％,(93. 7 ± 2. 7)％ and (96. 2 ± 2. 2)％,and the differences were all statistically significant(all P < 0. 05),respectively. Compared with other patients,the one with low expression of both E2F3a and CASP8AP2 had a poorer prognosis. In addition to MLL rear-rangements and minimal residual disease level at the end of remission induction,low expression of both E2F3a and CASP8AP2 remained as independent prognostic factors. Conclusion Low expressions of E2F3a and CASP8AP2 pre-dict poor prognosis in pediatric ALL. Combined assessment of E2F3a and CASP8AP2 expression could predict poor prognosis and relapse more accurately.