The Relationship Between DANCE And The Change Of Cell Metabolism Mode

Objective:Our goals were to study the change of metabolism mode in cells with differentintracellular densities by determination of mRNA and protein levels, and to find outthe relationship between DANCE and the change of metabolism mode in the cells.Methods:1) K562cells were divided into two groups, control group, coultured normally, andtest group, incubated with all-transretinoic acid in the coulture medium. The cellswere collected in48hrs after the drug treatment.2) Flowcytometric analysis was employed for apoptosis determination of K562cellsin bands S40and S50treated with VA, and those in bands S50and S60coulturednormally.3) Differential gene expressions between S40and S50of K562cells incubated withVA were determined by Illumina Cluster Station and Illumina HiSeq2000.The gene expression profileswere determined as mRNA levels in the cells.4) Real-time PCR and Western Blot were used to determine the mRNA and proteinexpression levels in K562cells of the separated bands.5) White blood cells (WBC) from a diabetic patient were separated by DSGC intotwo bands, S50and S60respectively.6) Real-time PCR and Western Blot were used to determine the levels of mRNA andprotein of some inflammation related genes, and the differences between thebands S60and S50of the WBC. Results:1) Normally coultured K562cells were separatedby DSGC into two bands, S60andS50.However, the K562cells incubated with all-trans retinoic acid wereseparated by DSGC into two different bands, S40and S50.2) Flowcytometric analysis showed that more cells were undergoing cellularnecrosis with decrease of the intracellular density.3) The gene expression profiles of VA incubated K562cells of S40and S50showeda significant difference in mRNA levels between the cells of the two bands.4) The results of Real-time PCR and Western Blot were found to be consistent withthe expression profilesdetermined by Illumina Cluster Station and IlluminaHiSeq2000.5) The expressions of CD36and IL-6inthe diabetic WBC were found to besignificantly different from each other between the bands S50and S60separatedby DSGC.Conclusions:DANCE reaction of K562cells and diabetic peripheral white blood cells wassignificantly accompanied by the change of gene expressions in the cells, suggestingthat DANCE was tightly related to the change of metabolism mode in the cells.