RESUMO
OBJECTIVE:To study the inhibi tory effects of genistein on the growth of human nasopharyngeal carcinoma. CNE 1 cells and predict its potential target. METHODS :CCK-8 method was used to test the effects of 0(blank control ),12.5,25,50, 100,150 µmol/L genistein on the proliferation of CNE 1 cells after treated for 24,48,72 h. Flow cytometry was carried out to detect the effects of 0(blank control ),15,30,60 µmol/L genistein on the cell cycle and ap optosis of CNE 1 cells after treated for 24 h. Scratch test was used to investigate the effects of 0(blank control ), 10, 20, 30 µmol/L genistein on themigration ability of CNE 1 cells after treated for 24 h. High (No.18210156) throughput sequencing was conducted to discover the differential genes in CNE 1 cells after treated with 0(blankcontrol),30 µmol/L genistein for 24 h. RT-qPCR assay was adopted to verify the mRNA expression of related differential genes in above trials. RESULTS : Compared with blank control,12.5,25,50,100,150 µmol/L genistein sho wed significant inhibitory effect on the proliferation of CNE 1 cells(P< 0.01),in a concentration- time-effect manner ;15,30 µmol/L genistein could arrest CNE 1 cell cycle at G 0/G1 stage(P<0.05 or P< 0.01);30,60 µmol/L could arrest CNE 1 cell cycle at G 2/M stage and promoted cell apoptosis (P<0.05 or P<0.01). 10,20,30 µmol/L genistein could significantly inhibit the migration ability of CNE 1 cells(padj<0.01). High throughput sequencing revealed a total of 2 271 differentialgenes(P<0.05),1 154 of which were up-regulated while 1 117 of which were down-regulated ;8 potential target genes ,including p53,p21,STC2,FGF2,CDK6,CYCLIN D ,PI3K,AKT,were screened by cell experiment. After validated by RT-qPCR assay ,mRNA expression of p53,p21,STC2,FGF2,CDK6,CYCLIN D and AKT were significantly down-regulated(P<0.05),which consistent with the sequencing results. CONCLUSIONS :Genistein can effectively inhibit the growth of human nasopharyngeal carcinoma CNE 1 cells,the mechanism of which may associated with inhibiting the expression of mutant gene p53,restoring the function of wild-type P 53 protein and inhibiting the activity of PI 3K/Akt pathway.
RESUMO
This study was purposed to confirm the practical efficacy of reducing indicating germs suspended in plasma by riboflavin and photosensitized inactivation and to evaluate its influence on activation of apheresis platelet concentrates. The synergistic effects of riboflavin combined with ultraviolet irradiation on inactivation of germs were investigated by using Escherichia Coli (E. coli) and Staphylococcus Aureus (S. aureus) as Gram⁻ and Gram(+) indicating germs, respectively. The activation status of apheresis-platelet concentrates treated with riboflavin combined with ultraviolet irradiation was detected by flow cytometry. The results showed that when 50 μmol/L of riboflavin was combined with 6.2 J/ml of ultraviolet irradiation, the T/E ratios reached 1.42 for E. coli and 1.68 for S. Aureus, and reduction of E. Coli and S. Aureus were 3.87 Logs and 3.82 Logs respectively; the CD62p expression level on germ-inactivated platelets stored at 22 degrees C for 0 and 5 days were 4.92% and 36.18% respectively, which slightly increased as compared with controls (3.94% and 32.03)% (p < 0.05). It is concluded that combination of riboflavin with ultraviolet irradiation displays well synergistic effects which can reduce E. Coli and S. Aureus counts, but no significantly influence on platelets. The partial activation of liquid platelets mainly presents metabolism damage during storage, which is found at an acceptable level.