Effect And Mechanism Of MiR-106b On Antithyroid Drug Induced Agranulocytosis

Objective : To observe the biological effects of mi R-106 b on HL-60 cells.Methods: HL-60 cells were cultured with medium 1640 and transfected the cells with mi R-106 b control/mimic/inhibitor.Then the expression level of mi R-106 b was detected by using quantificational real-time polymerase chain reaction(q RT- PCR). HL-60 cells were collected after 24 h,48h,72 h separately and measured its proliferation through MTT.48 h and 72 h after transfection, Annexin V/PI’staining under the fluorescence microscope and flow cytometry were used to observed cell apotosis.Result: Compared with the control group while q RT-PCR testing, the level of mi R-106 b was significantly decreased in mi R-106 b inhibitor group(p﹤0.05). After 48 h and 72 h, the HL-60 cells proliferation of inhibitor group was significantly suppressed than control group, while there was no obvious difference between mimic and control groups. The apoptosis rate of inhibitor group was highly increased than mimic and control groups.Conclusion: Mi R-106 b can promote neutrophil proliferation and inhibit apoptosis.Objective: To study the proapoptotic mechanism of mi R-106 b in the development of angranulocytosis. Methods: The bioinformation of mi R-106 b such as mature sequence, the target genes of related with cells apoptosis, binding sites of 3’UTR and targeting free energy were analyzed through online bioinformatics. We identified the target gene by dual-luciferase activity test. RT-PCR and Western-blot were used to detect the expression of m RNA and protein expression level of the target genes, and the change of related downstream proteins.Result: the software of bioinformatic analysis revealed a potential binding site for mi R-106 b within PTEN3’UTR, mi R-106 b got a high context score and low binding free energy of binding to PTEN3’UTR.Dual-luciferase activity cotransfected with mi R-106 b mimic and wild type recombinant plasmid were down-regulated in 293 T cells. Compared with mi R-106 b control by RT-PCR and Western-blot, the level of PTEN m RNA and PTEN protein was significantly increased in mi R-106 b inhibitor group. The protein level of p-Akt was downregulated while BAD and caspase-3 was upregulated according to the increase of PTEN protein.Conclusion: PTEN was confirmed as a target gene of mi R-106 b, mi R-106 b may contribute togranulocyte apoptosis by regulating PTEN.