Study On RhIL-13 Inducing Of The Differentiation Of HEL Cells And Its Mechanism

AIM: It is to investigate rhIL-13 induce the expression of proto-oncogene c-fos and its effects on the differentiation of HEL cells,partly clarify the signal transduction mechanism of IL-13. METHODS: 1.Cell culture: HEL cells were cultured in RPMI 1640 supplemented with 10% calf serum at 37℃with 5% CO₂ . Before the treatment of rhIL-13 ,HEL cells were grown to a density of 5×10⁸¹Ã—10⁹ /L and starved without serum for 20 hours . 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 some genes including that: (1)the expression of IL-13 receptor α1 (IL-13Rα1) mRNA in HEL cells incubated with rhIL-13 for 0,4,24 ,48 hours; (2)the expression of c-fos mRNA in HEL cells incubated with rhIL-13 for 0,15,30,60,120 minutes; (3)the expression of GPA( glycophorin A),GP(glycoprotein)â…¡b,vWF(von Willebrand Factor) mRNA in HEL cells incubated with or without rhIL-13 for 5 days. 4.Western Blotting: After HEL cells serum-starved for 20h, HEL cells were incubated with rhIL-13 for 0,15,30,60,120 minutes and RIPA Buffer was used for the lysis of HEL cells. The expression of c-Fos protein was observed by Western Blotting after the isolation of protein. RESULTS: 1.The expression of IL-13Rα1 mRNA is detected in HEL cells. 2. RhIL-13 induces the expression of c-fos mRNA in 15 min (P<0.05), reaches a peak in 30 min (P<0.01) and attenuates after 60 min. 3.RhIL-13 up-regulates the expression of GPIIb,GPA,vWF mRNA in HEL cells incubated with rhIL-13 for 5 days (P<0.05). 4. RhIL-13 enhances the expression of c-Fos protein in 30 min (P<0.05), reaches a peak in 60 min (P<0.01) and attenuates after 120 min. CONCLUSION: 1. HEL cells express IL-13Rα1 mRNA. 2. RhIL-13 induces the expression of c-fos mRNA and protein. 3. RhIL-13 up-regulates the expression of GPIIb,GPA ,vWF mRNA in HEL cells.