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Studies on Relationship between EMMPRIN Expression and Proliferation, Apotosis of Glioblastm Cell

Author: XuLiMin
Tutor: LuYongJian
School: Guangzhou Medical College
Course: Neurosurgery
Keywords: EMMPRIN siRNA U251 Cell Glioma
CLC: R739.41
Type: Master's thesis
Year: 2011
Downloads: 11
Quote: 0
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Abstract

The research background glioma is the most common primary central nervous system malignancies, accounting for about 44% of intracranial tumors. Due to the invasive growth characteristics of the tumor, surgery is difficult to accurately determine the boundaries of the tumor tissue and normal brain tissue. The one hand, due to the unnecessary expansion of the surgical area greatly increase patient residue, the probability of death; the other hand, even in a state-of-the-art surgical microscope assisted tumor tissue to the naked eye under total removal, but there are still about 104 to 105 The remnants of strong proliferative ability of tumor cells, and this is a glioma recurrence basis. Also have little effect due to the presence of the blood-brain barrier, resulting in limited existing anticancer drugs and normal brain tissue irreversible side effects. Related statistics show that the average survival period after the comprehensive treatment of malignant glioblastoma cell tumor is still less than 12 months, the 2-year survival rate was less than 10%. Recognized in glioma treatment around the world are the problem. That glioma etiology is not clear is the most important reason that its difficult to cure. The present study confirmed the occurrence of glioma development is a complex process involving multiple genes, a multi-stage, including the activation of oncogenes and tumor suppressor gene inactivation. Therefore, to reveal the molecular genetic mechanisms of glioma is of great significance to clarify the the glioma pathogenesis of reason and guidance in the genetic basis for the treatment of glioma. Human EMMPRIN gene is located on 19q13.3, about 60kb, contains seven exons and six introns, encoding a protein of 248 amino acids. The Sameshima study found that EMMPRIN mRNA expression was significantly lower than in normal brain tissue in glioma cells, the mRNA and protein expression levels with tumor grade and a positive correlation. However, inhibition of EMMPRIN gene expression in glioma cells, proliferation and apoptosis of glioma cells the impact has not been reported to the subject focuses on a the EMMPRIN gene on the proliferation and apoptosis of glioma cells impact, designed to provide the basis for the gene therapy of gliomas. Objective To study the inhibition of EMMPRIN gene expression in glioma cell proliferation and apoptosis of glioma cells. Research Methods U251 cell in a 37 ° C, 5% (v / v) CO2, saturated humidity condition in a culture medium containing 10% FBS, 100U/ml penicillin and 0.1mg/ml streptomycin in DMEM in culture. EMMPRIN-siRNA interference fragment by Shanghai GenePharma Limited completed the design and synthesis. EMMPRIN mRNA expression was detected by RT-PCR, GAPDH as a loading control. EMMPRIN expression by Western blot, Tubulin as a loading control. EMMPRIN-siRNA interference segment of the MTT assay transfection inhibits EMMPRIN expression on U251 cell proliferation. EMMPRIN-siRNA interference fragment of flow cytometry transfection inhibits EMMPRIN expression on U251 apoptosis and cell cycle distribution change. SPSS13.0 statistical package for statistical analysis. EMMPRIN-siRNA interference fragment results U251 cell transfection significantly inhibited U251 cells EMMPRIN gene expression at the mRNA and protein levels. EMMPRIN-siRNA interference segment of the MTT assay is U251 cell transfection of U251 cell proliferation was significantly inhibited. EMMPRIN-siRNA interference fragment U251 cell transfection of U251 cell apoptosis increased significantly. EMMPRIN-siRNA interference fragment U251 cell transfection of U251 cell cycle distribution of a significant change in a significant reduction in the proliferative phase cells. Conclusion EMMPRIN gene-specific siRNA can inhibit glioma U251 cell line proliferation and promote apoptosis.

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