The Study On The Mechanisms Of SIL-1RAP In Low-Grade Glioma

Objective Glioma is the most common human tumors of the central nervous system(CNS). Wherein,40%to60%of the brain tumors have a wide range of aggressivetendency from low level to a high level of conversion. Some related studies show thatlow-grade gliomas (LGGs) account for33%of all CNS, then in adults LGGs account for25%and35%of childhood. Although, the pathological grade and the localization inLGGs patients are the same (except the brain stem), the clinical progression andprognosis are quite different between adults and children. Adult patients with low-gradeastrocytomas (except cell astrocytoma) are much easier to maligne to the higher degree ofmalignancy malignant glioblastoma, but the probability of malignancy is lower inchildren, and the prognosis is also good. The different prognosis of LGGs betweenchildren and adults, suggest different biological characteristics of both. Our previousstudies showed that sIL-1RAP significantly differentially expressed in LGGs betweenchildren and adults. Moreover, we also found that sIL-1RAP and STAT3co-localizationin the nucleus in U251, but existing studies indicate that sIL-1RAP is not localized in thenucleus. So, researches on the mechanism of their nuclear co-localization can help us toreveal the molecular mechanism of sIL-1RAP in LGGs with different prognostic betweenadults and children.Methods (1) The total RNAs were extracted from fresh surgical specimens and themRNA levels of sIL-1RAP were detected by RT-PCR.(2) At the same time, western blotwere used to detect the protein levels of sIL-1RAP.(3) The glioma cell line U251wastransfected with the siRNA of STAT3. Then the protein levels of STAT3were detectedby western blot.(4) Later, we extracted the RNAs from U251cells, then, detectedSTAT3by RT-PCR and qRT-PCR in the levels of mRNA.(5) We transfected U251cellswith both the siRNA of STAT3and the plasmid of sIL-1RAP. Then immunofluorescencewas used to detect the levels of sIL-1RAP in the nucleus when we suppressed the proteinlevels of STAT3.Results As taking RNA interference in our study, we also use Immunofluorescent tecn-niques to verify the effect of STAT3during the sILRRAP into the nucleus, and we fouund that inhibited STAT3could not affect sIL-1RAP into nucleus.Conclusion The reason of sIL-1RAP can not into nucleus may the poorer inhibition of STAT3or there may have other compensatory mechanism to assist sIL-1RAP into thenucleus. As sIL-1RAP into nucleus has significant effect on glioma occurrence anddevelopment, so we can use sIL-1RAP into the nucleus as a potential drug-target for thetreatment of glioma and we will continue to the depth study of inhibition sIL-1RAPinto the nucleus.