The Preliminary Study Of The Signal Transduction Of P38MAPK On The Cell Cycle Of Rat Glioma Cells C6

MAPK cascade is an important signal transducer from extracellular to nuclear. The cascade can activate the intracellular proteins, which take part in signal transduction and affect many physilogical and pathological processes. p38MAPK is a new kind of MAPK subtribe found in recent years, which takes part in cell survival, devision and apoptosis, etc, TNF- a can activate p38MAPK pathway, but the effect in the apoptosis after its activation remains unknown. In the same time, p38MAPK can upregulate the expression of TNF-a and induce apoptosis. Glioma is the most common tumor in the central nervous system. The characters such as infiltrative growth, strong invasion, metastasis in early stage and easy recurrence, are related to the disorder of signal transduction in tumor cells probably. The effect of p38MAPK pathway on the growth, development of glioma and its regulation of the cell cycle of glioma together with TNF- a are not clear.AIM (1) To study the effect of p38MAPK on morphological change and cell cycle of C6 cells. (2) To study the effect of TNF- a on the proliferation of C6 cells and the expression of p38MAPK.(3) To study the mechanisms of the effect of p38MAPK on the cell cycle of C6 cells preliminarily.METHODS (1) p38MAPK was transfected into C6 cells by lipofectin. Expression of p38MAPK was detected by immunocytochemistry and Western-blot. HE. Staining, transmission electron microscopy(TEM) and flow cytometry were adopted to measure the cell morphology and cell cycle. (2) The effect of TNF- a on the proliferation of C6 cells was determined by MTT assay. The apoptosis was detected by TEM and flow cytometry. The expression of p38MAPK was detected by immunocytochemistry and Western-blot. (3) The concentration of sTNF-awas detected by ELISA, the concentrations of membrane TNF- a and TNFRI were detected by flow cytometry.RESULTS (1) p38MAPK expressed in C6 cells after transfection, with cell morphology changing and apoptotic cells emerging. (2) The inhibitory rate of TNF- a on C6 cells was 43.75%. Apoptotic cells were detected by TEM and apoptotic rate was 37.5% by flow cytometry. p38MAPK positive signals were detected by immunocytochemistry and Western-blot. (3) The concentration of sTNF- a increased obviously. The concentration of member TNF- a had no obvious change, and that of TNFRI increased.CONCLUSION (1) The apoptosis of C6 cells could be induced by p38MAPK transfection. (2) The apoptosis of C6 cells and expression of p38MAPK could be induced by TNF-a. The activation of p38MAPK was related to the apoptosis of C6 cells probably. (3) The increase of sTNF- a and membrane TNFRI may be one of the reasons why p38MAPK transfection induced the apoptosis of C6 cells.