Correlation Analysis Of IGFBP-3with Malignant Glioma

Objective: As the most common malignant intracranial tumor, glioma isaccounting for40-50%of all brain tumors. Accordance with the2000WHOstandard pathological grading standards, glioblastoma can be divided inPilocytic astrocytoma (grade Ⅰ), dispersive astrocytoma (grade II),anaplastic astrocytoma (grade III) and neuroblastoma (grade IV). As reported,annual incidence generally3-6/100,000of glioma, showed an increasingtrend year by year, and the annual growth rate was1.2%. Glioma is still one ofthe worst prognosis tumors. Thus, the study is an important topic todevelopment, the mechanism of invasion and recurrence of glioma,development of new anticancer drugs, reduce the threat to human life of braintumor.IGFBP-3which is one of the members of the IGFBPs family is the bloodof IGF binding protein to bind and transport of IGF-I and modulate itsactivity, thereby playing an important role in cell growth. Recent in vitrostudies show that IGFBP-3, an important regulator, play a role on tumor cellsurvival through non-dependent IGF mechanisms.Multiple signalingpathways within the cell, such as extracellular signal-regulated kinase (ERK),nuclear transcription factor kappa B (the Nuclear factor-kappa b, of NF-κB),Smad2/3, etc involved in the incidence, development, regulation, invasionand recurrence of tumors, especially ERK signal transduction pathway plays avital role in regulating proliferation,survival and apoptosis of glioma cells.IGFBP-3can inhibit the activation of the ERK signal transduction pathwayand play a pro-apoptotic anti-proliferation, but the specific mechanism is stillunclear.This study detected expression of IGFBP-3in malignant glioma byimmunohistochemical staining method and Q-PCR. We want to explain themolecular mechanism of apoptosis regulation of IGFBP-3in malignant glioma cells and improve the IGFBP-3non-dependent IGF-I through theERK signaling pathway to inhibit cell proliferation and promote apoptosistheory to reveal the mechanism of recurrent malignant glioma. We can find anew target of the regulation of apoptosis of malignant glioma and provide newideas and methods for clinical treatment of malignant glioma.Methods:1Collection of clinical specimens: the last three years, we collectedpathological grade II-IV grade glioma specimens of34cases (pathologicgrade based on WHO2000grading standards) in the Second Hospital of HebeiMedical University, including II grade astrocytoma glioma specimens of11cases, III grade astrocytic glioma specimens of14cases, IV grade astrocyticglioma specimens9cases, while taking the16cases of normal brain tissues(control group).2Detection and analysis of clinical specimens: Expression of IGFBP-3wasdetected quantitative on mRNA levels of in all levels of glioma and normaltissues by Q–PCR. By immunohistochemical methods detected theexpression of IGFBP-3on protein levels.3Statistical analysis: Use the spss17.0statistical software to analysisexperimental datas.Results:1After4%formaldehyde solution fixed glioma specimens and normal braintissues, expression of IGFBP-3was analyzed by immunohistochemicalmethods, and the positive cells showed brown. The application spss17.0statistical analysis showed that expression of IGFBP-3in human gliomas werehigher than those in normal brain tissues, this difference was statisticallysignificant, P values less than0.05. Expression of IGFBP-3in gliomas of alllevels, there were differences, and P values was less than0.05, this differencewas statistically significant. These results showed,that expression of IGFBP-3in human glioma compared with normal brain tissue are apparently increasedon the protein level, and this difference related with increasing pathologicalgrade of gliomas. 2After using RNA extraction kit to extract total RNA from the glioma cellsand normal brain tissue cells, UV spectrophotometric determination of RNAconcentration (the A260/A280>1.8) and1%agarose gel electrophoresis(including EB0.5mg/l) observed in the28s and18s RNA two clearbelt.IGFBP-3as a targeted gene will be amplified by Q-PCR technology andtagged fluorescent amplified enzyme MaximaTMSYBR Green/ROX Q-PCRMaster Mix (2×). Application spss17.0statistical software for statisticalanalysis results showed that expression of IGFBP-3in glioma cell groupcompared with normal brain tissue group was significantly increased, and thisdifference was statistically significant, P values were less than0.05,expression of IGFBP-3in grade II, III, IV glioma cells groups showed anincreasing trend, and P values were less than0.05, the difference wasstatistically significant. The results demonstrated, expression of IGFBP-3inhuman gliomas compared with normal brain tissue was significantly increasedon the genetic level, and increased gradually with the malignant progression ofgliomas.Conclusion:1Expression of IGFBP-3increased significantly in malignant gliomacomparing with the normal craniocerebral tissues.2Expression of IGFBP-3were different in diverse grade gliomas, showing anincreasing trend with the increased levels of malignancy. IGFBP-3is closelyrelated with the progress of glioma.3IGFBP-3can play a role of inhibiting the activation of the ERK signaltransduction pathway, promoting tumor cell apoptosis and inhibitingproliferation cell by dependent or non-dependent IGF, and may provide newtargets and new ideas to the clinical treatment of glioma.