Objective To explore the inhibitory effect of anti-miRNA-214 oligonucleotide on leukemia U937 cells.Methods U937 cells were transfected with anit-miRNA-214 oligonucleotide,cell viability was analyzed by MTT assay.Apoptosis was detected by flow cytometry.The expression of miRNA214 in the U937 cells were measured by real-time PCR.Results The MTT result showed that the growth of U937 cells treated with AMO-miRNA-214 in 24 h,48 h,72 h was obviously inhibited (0.812±0.001,0.770±0.002,0.541±0.001),compared with those in control groups (randomized control 1.011±0.002,1.112±0.003,1.111±0.003,blank control 1.112±0.001,1.023±0.001,1.101±0.001),the differences had statistical significance (F =2.782,3.659,2.735,P =0.021,0.018,0.036).The flow cytometry results showed that the apoptosis detected in AMO-miRNA-214 group at 48 h,72 h (15.12±0.02,19.14±0.01) had significant differences compared with those in control groups (randomized control 2.04±0.02,2.45±0.03,blank control 1.19±0.02,2.02±0.01) (F =3.683,3.762,P =0.013,0.015).The expression of miRNA-214 was downregulated significantly in U937 cells after treated with oligonucleotide (31.1±0.2) compared with those in control groups (randomized control 25.8±0.1,blank control 25.6±0.2) (P ＜ 0.05).Conclusion Targeted inhibition of miRNA-214 with oligonucleotide can suppress U937 cells growth and induce apoptosis.

To use Agrobacterium rhizogenes-induced hairy roots to create new germplasm of Dianthus caryophyllus, we transformed D. caryophyllus with A. rhizogenes by leaf disc for plant regeneration from hairy roots. The white hairy roots could be induced from the basal surface of leaf explants of D. caryophyllus 12 days after inoculation with A. rhizogenes ATCC15834. The percentage of the rooting leaf explants was about 90% 21 days after inoculation. The hairy roots could grow rapidly and autonomously in liquid or solid phytohormone-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and silica gel thin-layer chromatography of opines from D. caryophyllus hairy roots. Hairy roots could form light green callus after cultured on MS+6-BA 1.0-3.0 mg/L + NAA 0.1-0.2 mg/L for 15 days. The optimum medium for adventitious shoots formation was MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L, where the rate of adventitious shoot induction was 100% after cultured for 6 weeks. The mean number of adventitious shoot per callus was 30-40. The adventitious shoots can form roots when cultured on phytohormone-free 1/2 MS or 1/2 MS +0.5 mg/L NAA for 10 days. When the rooted plantlets transplanted in the substrate mixed with perlite sand and peat (volume ratio of 1:2), the survival rate was above 95%.
Agrobacterium ; Chromatography, Thin Layer ; Culture Media ; Dianthus ; growth & development ; Plant Growth Regulators ; Plant Leaves ; Plant Roots ; growth & development ; Plants, Genetically Modified ; Rhizobium ; Tissue Culture Techniques ; Transformation, Genetic