Discussing The Relationship Between HOXA5 Expression And Cell Cycle Or Apoptosis Under The ATRA Intervention In K562 Cells

Objective:This paper aims to analyze the expression of Homeobox genes,HOXA5 and its relation to the cell cycle, apoptosis in human myeloid leukemia cell line K562 cells intervened by All-Trans Retinoic Acid to make an initial evaluation of its effects and significance in myeloid leukemia in the cytogenesis and development process, and the correlation between them. Methods:1. Treat human myeloid leukemia K562 cells as experimental models by tumor cells in vitro cultivation technology.2. Cell proliferation- toxicity experiment(Cell Counting Kit-8 CCK-8):To detect proliferation inhibition capacity of K562 cell with different concentrations of ATRA intervention 24 h, 48 h, 72 h, and 96 h to determine the best density ATRA intervention in K562 cells.3. Group of Experiments: The logarithmic phase cells(about 105 / ml), K562 cells were randomly divided into ATRA intervention group(experimental group) and control group. Experimental group set up three time subgroups, with 10.0 umol/L ATRA intervention 24 h, 48 h, and 72h; The control group used medium instead of ATRA and the rest was same as ATRA intervention group.4. Flow cytometry detection: To Measure cell apoptosis situation under the intervention of 10.0 umol/L ATRA on K562 cells after 24 h, 48 h, and 72 h.5. Flow cytometer detection: Detect and analysis of 10.0 umol/L ATRA intervention in K562 cells after different time, compared with the control group corresponding changes in the cell cycle distribution.6. Real-time fluorescent quantitative PCR(PT-PCR) technique detection: Measure 10.0 umol/L ATRA intervention in K562 cells after 24 h, 48 h, and 72 h HOXA5mRNA expression in cells, and analyze the expression of HOXA5 mRNA and its relationship with apoptosis.7. Western blot detection: Measure 10.0 umol/L ATRA intervention in K562 cells after 24 h, 48 h, and 72 h HOXA5 protein expression in cells, and analyzes the relations between HOXA5 protein expression and apoptosis.8. Statistical analysis: Dispose the experimental results by statistical software SPSS 17.0, with P<0.05 for the difference was statistically significant. Correlation analysis between the two variables were the test of Spearman rank correlation analysis, a=0.05 for the test of significance level.1. CCK-8 result display: ATRA could inhibit the proliferation of K562 cells, the experimental group compared with control group, the OD value of the difference was statistically significant(P<0.05). When concentration of ATRA intervention was 10.0 umol/L, each cell at different time points had the best growth trend.2. Flow cytometry to detect cell apoptosis rate result display: Experimental cell apoptosis rate was significantly higher than control group(P<0.05), and the experimental group with drug intervention in the extension of time, the cell apoptosis rate also gradually increased was higher than normal control group(P<0.05), and longer duration of intervention HOXA5 protein expression quantity was increased(P<0.05).6. Relation between HOXA5 expression and apoptosis display: The expression of HOXA5 mRNA,protein and apoptosis in K562 cells showed that both were positively correlated(P<0.05) by the Spearman rank correlation analysis.7. Relation between HOXA5 expression and cell cycle display: The expression of HOXA5 mRNA,protein and cell cycle in K562 cells showed Cell cycle G0 / G1 phase increased percentage were positively correlated, and negatively correlated with S phase lower percentage. Conclusion:1. With intervention of ATRA,in human myeloid leukemia cell line K562 cell, HOXA5 mRNA and protein expression of the quantity increased in time.2. ATRA could inhibit cell proliferation and promote apoptosis of K562 cells by raising HOXA5 mRNA and protein expression.