The Expression And Effect Of Cdk11^p58 In Schwann Cells Under Inflammation

cyclin D3 molecules in rat RSC96 cells models of inflammation, and investigate their function in the process of the biological behaviour of Schwann cells. Methods1. The model was prepared in rat RSC96 cells by LPS. The cell content of PCNA and activated caspase-3 protein were assayed by western blot.2. Western blot and RT-PCR were used to detect the changes of CDK11 and cyclin D3 protein expression in the LPS stimulation of Schwann cells, the distribution and interaction of CDK11 and cyclin D3 were investigated by Immunofluorescen,Cellular Fractionation and Immunoprecipitation.3. Transfecting the cyclin D3 siRNA vector and HA-p58 overexpression vector into SCs in order to investe the distribution of CDK11^p58 and the influence in Schwann cells proliferation and apoptosis.4. Cultured Schwann cells were stimulated by LPS. The extracellular signal-regulated kinase (ERK) 1/2 inhibitor, U0126, P38 inhibitor, SB202190, and SAPK/JNK specific inhibitor, SP600125 were used to analysis the signal pathway of LPS-induced CDK11^p58 expression.Results1. In the model of LPS induce SCs, the results indicated that LPS can induce caspase-3 activate and PCNA expression in a dose and time-independent.2. Cultured Schwann cells were stimulated by LPS. The expression pattern of CDK11^p58 and cyclin D3 in the activate Schwann cells was in a dose and time-dependent increasing. In the normal Schwann cells, CDK11^p58 and cyclin D3 express in nucleus and cytoplasm. While by the stimulating of LPS, the two proteins colocalized with each other in the activated Schwann cells, and mainly in nuclear regions. By Immunoprecipitation, we found that CDK11^p58 and cyclin D3 interacted with each other in the Schwann cells, and mainly in nuclear regions. LPS may enhance this proces.3. After transfecting HA-p58 vector, we found that overexpression of CDK11^p58 repressed Schwann cell proliferation and induce Schwann cell apoptosis. After transfecting cyclin D3 siRNA vector, we found that Silence cyclin D3 decreased CDK11^p58 nuclear translocation in SCs and decreased Schwann cell apoptosis.4. The extracellular signal-regulated kinase (ERK) 1/2 inhibitor, U0126, P38 inhibitor, SB202190, and SAPK/JNK specific inhibitor, SP600125 were used to analysis the signal pathway of LPS-induced CDK11^p58 expression. We found that LPS regulated CDK11^p58 expression in Schwann cells via the activation of MAPKs, especially SAPK/JNK pathways, and inhibitors specific to MAPK subgroups could block CDK11^p58 expression in SCs and block its activity of kinase. While ERK1/2 had no influence on Schwann cells, and p38 had no obviously influence.After transfecting HA-p58 vector, we found that overexpression of CDK11^p58 repressed the expression of BCL-2, and LPS may enhance this process. But SAPK/JNK specific inhibitor, SP600125 may obviously inversion this process.Conclusions1. In the normal Schwann cells, CDK11^p58 and cyclin D3 express in nucleus and cytoplasm. While by the stimulating of LPS, the expression of the two proteins increased, and mainly in nuclear regions. These dataes indicate that CDK11^p58 is an inflammatory response molecule participating in the inflammatory process of never systerm.2. After transfecting HA-p58 vector, the overexpression of CDK11^p58 can induce caspase-3 activate and depress PCNA expression indicate that CDK11^p58 may repress Schwann cell proliferation and induce Schwann cell apoptosis. After transfecting cyclin D3 siRNA vector, the silence cyclin D3 decreased CDK11^p58 nuclear translocation in SCs and decreased the expression of activated caspase-3. These data indicate that cyclin D3 may regulate the expression of CDK11^p58 and its activity of kinase.3. Cultured Schwann cells were stimulated by LPS, the expression pattern of CDK11^p58 and cyclin D3 in the activate Schwann cells was in a dose and time-dependent increasing, and this process can repress the expression of BCL-2. These effects are regulated by SAPK/JNK signal pathway.