Application of E3 ubiquitin ligase CHIP in gliomatosis cerebri disease
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A ubiquitin ligase, glioma technology, applied in the application field of E3 ubiquitin ligase CHIP in glioma disease, can solve problems such as proliferation and invasion
Inactive Publication Date: 2010-12-29
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Previous studies have suggested that CHIP is highly expressed in brain tissue, but its specific expression localization and its relationship with the development, proliferation and invasion of glioma have not been reported internationally.
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Embodiment 1
[0042] Expression of CHIP in glioma tissues and cell lines and its relationship with pathological grade
[0043] In this example, real-time quantitative RT-PCR, immunohistochemistry, immunofluorescence and other methods were used to study the expression of CHIP in glioma tissue cell lines and its relationship with pathological grades.
[0044] Materials and Methods
[0045] 1. Experimental materials
[0046] (1) Tissue specimens
[0047] Selected 20 cases of brain glioma tissue from Shanghai Changzheng Hospital from 2002 to 2009, which were surgically resected, confirmed by pathology, and embedded in paraffin, including 10 cases of low-grade glioma (5 cases of pilocytic astrocytoma, diffuse astrocytoma 5 cases of tumor), 10 cases of high-grade glioma (5 cases of anaplastic astrocytoma, 5 cases of glioblastoma). Five cases of paraffin-embedded normal brain tissue. All pathological diagnoses and grading were determined independently by two pathologists.
[0048] Both U251 ...
Embodiment 2
[0233] Construction and identification of RNA interference and overexpression lentiviral vector targeting CHIP
[0234]Example 1 has proved that: in normal brain tissue, CHIP is mainly expressed in the cytoplasm of neurons, but not in normal glial cells; in glioma tissues, CHIP is mainly expressed in the cytoplasm of tumor cells , co-localized with the marker protein GFAP of astrocytes, and its expression increased with the increase of the level, suggesting that it may play a role in the development, proliferation and invasion of glioma. However, this finding is in contrast to the expression of CHIP in other tumors. In order to further study the biological function of CHIP at the cellular and animal levels, this example intends to construct a lentiviral RNA interference and overexpression vector targeting CHIP to further study the biological function of CHIP.
[0235] Materials and Methods
[0236] 1. Experimental reagents
[0237] PCR reagent primer(R&F)
[0238...
Embodiment 3
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Abstract
The invention relates to application of E3 ubiquitin ligase CHIP in a gliomatosis cerebri disease. In glioma tissues, CHIP is mainly expressed in endochylema of a tumor cell and is localized with labeled protein GFAP (Glial Fibrillary Acidic Protein) of an astroglia, and the expression increases along with the increment of levels; after a CHIP expression amount rises, mRNA (Messenger RNA) and protein expression of a Survivin increase; and after the CHIP expression amount reduces, the mRNA (Messenger RNA) and the protein expression of the Survivin decrease and the protein expression of beta-catenin increases. Through immunohistochemistry, the invention compares the expression conditions of the CHIP in gliomas at different levels and normal gliomatosis cells, clarifies effects and pathological mechanisms of the CHIP in the processes of glioma occurrence, development, proliferation and invasiveness and provides powerful means for the biological target treatment of the glioma in future.
Description
technical field [0001] The invention relates to the research field of the pathogenesis of brain glioma, and in particular relates to the application of an E3 ubiquitin ligase CHIP in brain glioma disease. Background technique [0002] Glioma, referred to as glioma, is a tumor that occurs in the neuroectoderm. There are two types of neuroectodermal tumors, one is formed by mesenchymal cells, called glioma; the other is formed by parenchymal cells, called neuronal tumors. Since the two types of tumors cannot be completely distinguished in terms of etiology and morphology, and gliomas originating from stromal cells are much more common than neuronal tumors originating from parenchymal cells, neuronal tumors include gliomas. Gliomas are collectively referred to as gliomas. [0003] Most of the gliomas develop slowly, and the time from the onset of symptoms to seeing a doctor usually takes several weeks to several months, and a few can reach several years. Highly malignant and...
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