Study On IL-13 Inducing Of STAT6 Phosphorylated In Dami And HEL Leukemia Cell Lines

Objective: rhIL-13 (recombinant human interleukin 13, rhIL-13) inducing the expression and activation of STAT6 (Signal transducers and activators of transcription) was investigated in Dami and HEL cells to elucidate the signal transduction mechanism of IL—13.Methods: 1.Cell culture: Dami and HEL cells were cultured in RPMI 1640 supplemented with 10% calf serum at 37°C with 5% CO₂. Before the treatment of rhIL-13 , Dami and HEL cells were grown to a density of 8×10⁸～1×10⁹/L and cultured for 20 hours in serum-free medium. 2. Isolation of total RNA: Total RNA was extracted from those cells incubated with or without rhIL-13 100μg/L respectively for various time points . The RNA quality was measured by means of RNA agar gel electrophoresis. 3. Reverse transcription polymerase chain reaction (RT-PCR) was used to observe the expression of IL-4 receptor α (IL-4Rα) mRNA in Dami and HEL cells incubated with rhIL-13 for 0, 4, 12 hours. 4. Western blot: After Dami and HEL cells were cultured for 20 hours in serum-free medium, cells were incubated with rhIL-13 for 0, 1, 2, 4, 8 hours and RIPA buffer was used for the lysis of cells. The expression of toal STAT6 and phosphorylated STAT6 (p-STAT6) was detected by Western blotafter the isolation of protein.Results:1. STAT6 are expressed in Dami cells. The phosphorylation of STAT6 is induced after 2 hours of rhIL-13 treatment and dephosphorylation after 4 hours in Dami cells.2. STAT6 are expressed in HEL cells. The phosphorylation of STAT6 is induced after 1 hours of rhIL-13 treatment and dephosphorylation...