RhIL-13 Mediates Expression Of Proto-oncogene C-fos And Its Effect On The Differentiation Of Dami Cells

AIM: It is to investigate that recombinant human interleukine-13(rhIL-13) mediates expression of proto-oncogene c-fos and its effect on the differentiation of megakaryocytic cell line, Dami cell, and provide the experiment and theory evidence for the clinical practice of IL-13. METHODS: 1.Effect of rhIL-13 on the growth of Dami cells was measured by means of Dami cell proliferation experiments including diagram of curves on the growth of Dami cells and MTT. 2. After 1×109/L Dami cells serum-starved for 20h, total RNA was extracted from those cells treated by rhIL-13 separately for 30 or 60 min or 5 generations. 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 some genes including that:(1)the expression of IL-13 receptorα1 chain(IL-13Rα1) in Dami cells;(2)the expression of c-fos gene in Dami cells treated for 30min or 60min by rhIL-13;(3)the expression of GPIIb and Glycophorin A(GPA) genes in Dami cells treated for 5 generations by rhIL-13. 4. After Dami cells serum-starved for 20h, Dami cells were treated by rhIL-13 for 30 min and RIPA Buffer was used for Dami cells lysis. The change of c-fos protein expression was observed by Western Blotting. RESULTS: 1.The expression of IL-13 Rα1 is detected in Dami cells by RT-PCR. 2.The c-fos mRNA expression were observed in rhIL-13 treated group(100ug/L),serum treated group(15%serum) and mixture treated group (100ug/LrhIL-13+15%serum) after Dami cells treated for 30 min,while c-fos mRNA expression disappears in rhIL-13 treated group and mixture treated group after 60 min except serum treated group.3.RhIL-13 up-regulates the expression of GPIIb, a megaKaryoc- ytic`s surface marker, in Dami cells treated by rhIL-13 for 5 generations. 4. RhIL-13 up-regulates the expression of GPA gene in Dami cells treated by rhIL-13 for 5 generations. 5. In contrast to the blank control group,the expression of c-fos protein is obviously enhanced in rhIL-13 treated group,serum treated group and mixture treated group after Dami cells treated for 30 min. 6.The result of MTT shows no dominant difference between the control group and rhIL-13 treated group,which indicates that rhIL-13 has no positive effect on the proliferation of Dami cells(P>0.05). CONCLUSION: 1. Dami cells express IL-13 receptor α1. 2.RhIL-13 induces c-fos mRNA and protein expressions in Dami cells. 3.RhIL-13 up-regulates the mRNA expressions of GPA and GPIIb.4. RhIL-13 hardly exerts dominantly positive effect on the proliferation of Dami cells.