Cabonylreductase 1(CBR1)expression Variability In Human Tongue Carcinoma Tissue And Cells And Regulation Mechanism

Objective:Using human tongue carcinoma tissues and eight cancer cells,we determined carbonyl reductase 1(CBR1)expression variability.Factors associated with CBR1 expression were also explored.Effect of over-expression of CBR1 on cell migration in A549 cells were studied.Methods:(1)Human tongue carcimoma tissues were collected.DNA,RNA,small RNA,and proteins were extracted from those tissues.CBR13’-UTR SNP rs9024 were determined by direct sequencing.Quantitative PCR were conducted to determine copy number of CBR1 mRNA,hsa-miR-574-5p and hsa-miR-921.Westen blot was conducted to evaluate protein expression of CBR1.Correlation between rs9024 genotype,copy number of CBR1 mRNA,hsa-miR-574-5p and hsa-mi R-921 were analyzed via SPSS.(2)CBR1 expression variability was evaluated in eight human cancer cells respectively: UM1、HepG-2、 SCC-15、MCF-7、 A549、K562、 DU145 and Hela.Cell scratch test was conducted to compare migration ability of those cancer cells.(3)CBR1pENTER expression plasmid was transiently transfected into A549 cells.Transwell cell test were conducted to evaluate CBR1 over-expression on A549 cell migration respectively.Results:(1)A total of 21 human tongue carcinoma tissues were collected from patients.CBR1 protein expression,copy number of CBR1 mRNA showed interindividual variability.CBR1 protein expression was positively correlated with copy number of CBR1mRNA(R2 = 0.711,p = 0.01).CBR1 rs9024 G/G genotype showed higher CBR1 protein expression than rs9024 G/A genotype(p = 0.04).Copy number of hsa-miR-574-5p and hsa-mi R-921 were not associated with CBR1 protein expression.(2)UM1、HepG-2、 SCC-15、MCF-7、 A549、K562、 DU145 and Hela cells all carried wild type CBR1 rs9024 genotype.CBR1 protein expression level varied by 45 fold among those cells.48 hours after cell scratch test,SCC15,DU15 and UM1 migrated faster than other cancer cells.(3)Over expression of CBR1 in A549 cells significantly decreased migration ability of A549 cells.ConclusionThere was great variability in CBR1 protein expression in human tongue carcinoma tissues,which was associated with CBR1 mRNA expression and CBR1 rs9024 genotype.CBR1 expression was not regulated by hsa-miR-574-5p and hsa-miR-921、 UM1、 HepG-2、 SCC-15、 MCF-7、A549、 K562、 DU145 cells showed variability in CBR1 expression level and cell migration.CBR1 expression in those cells was not associated with cell migration ability.Over expression of CBR1 may inhibit migration in A549 cells.