Composition for inhibiting glioma growth and application thereof

A glioma and composition technology, applied in the field of glioma growth-inhibiting compositions, can solve problems such as highly malignant phenotypes of GBM, and achieve colony formation inhibition, broad application prospects, and increased cell apoptosis rate Effect

Inactive Publication Date: 2014-05-14
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Amplification of EGFR and loss of PTEN lead to hyperactivity of PI3K/AKT signaling pathway, which

Method used

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  • Composition for inhibiting glioma growth and application thereof
  • Composition for inhibiting glioma growth and application thereof
  • Composition for inhibiting glioma growth and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, RNA interference recombinant expression vector construction

[0047] 1. Selection of RNA interference target sequence

[0048] 1) For the full-length cDNA sequence of the protein (referred to as protein B) shown in Sequence Listing 3 (referred to as Protein B) (as shown in Sequence Listing 4, that is, the PIK3CB gene), select the following two DNA sequences as the RNA interference suppression sequence 3 Indicates the target sequence for protein expression:

[0049] sh-1: position 1166-1184 of sequence 4 in the sequence listing (ie 5'-CCACTGGAATTTGATATTA-3');

[0050] sh-2: No. 439-457 of sequence 4 in the sequence listing (ie 5'-GGATCCTGAAGTAAATGAA-3');

[0051] The 17th to 3229th positions in the sequence listing sequence 4 are open reading frames, encoding the protein shown in the sequence listing sequence 3, which is a catalytic subunit p110 of PI3K beta subtype lipid kinase.

[0052] 2) For the full-length cDNA sequence (NCBI number: NM_002645, PIK3...

Embodiment 2

[0107] Embodiment 2, the recombinant expression vector construction of protein PTEN

[0108] Insert the coding gene (as shown in Sequence Listing Sequence 2) of protein PTEN (as shown in Sequence Listing Sequence 1) into BamH I and NotI of doxycycline-induced retroviral expression vector pRetro-on (BD Clontech Company) Between the sites, the recombinant expression vector PTEN / pRetro-on was obtained. It was confirmed by sequencing that the vector PTEN / pRetro-on was inserted between the BamH I and NotI sites of pRetro-on with the DNA molecule shown in Sequence 2 of the Sequence Listing.

Embodiment 3

[0109] Example 3, the influence of the expression of recovery protein PTEN co-inhibitor protein B on the phosphorylation level of AKT in recombinant glioma cells

[0110] Human U251 glioblastoma cell line (U251): the number in the cell bank of China Center for Type Culture Collection is 3115CNCB00312;

[0111] Human SHG-44 glioblastoma cell line (SHG-44): Cell Bank of the Chinese Academy of Sciences, catalog number TCHu48;

[0112] Both U251 and SHG-44 are glioma cell lines with PTEN deletion;

[0113] The culture conditions of the recombinant glioma cells of Examples 3-5 are as follows unless otherwise specified:

[0114] Medium: DMEM medium (Gibco, Cat. No. 12800-017), supplemented with 10% heat-inactivated fetal bovine serum (FBS), 4mM glutamine, 50units / ml penicillin and 50μg / ml streptomycin;

[0115] Culture conditions: 37°C, 5% CO 2 Incubator cultivation.

[0116] 1. Construction of stable recombinant glioma cell lines

[0117] The vector PTEN / pRetro-on obtained in ...

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Abstract

The invention discloses a composition for inhibiting glioma growth and an application thereof. The composition comprises a substance for improving the expression of protein (PTEN) as shown in sequence 1 of the sequence table, and a substance for inhibiting the expression of protein (B) as shown in sequence 3 of the sequence table. Experiments demonstrate that when the combination of recovering the expression of protein PTEN with inhibiting the expression of protein B is compared with only recovery of the expression of protein PTEN or only inhibition of expression of protein B, the AKT phosphorylation level of recombinant glioma cell lines is significantly decreased, cell proliferation and colony formation are significantly inhibited, and the proportion of cells stopping at the G0/G1 phase and the cell apoptosis rate are significantly increased; Glioma in transplanted mouse body has no increase in size at 20-48 days after treatment by recovering the expression of protein PTEN combined with inhibiting the expression of protein B, and the tumor weight is almost zero at the 48th day after the treatment. The invention provides a new and effective combination therapy scheme for glioma, and has wide application prospects.

Description

technical field [0001] The invention relates to a composition for inhibiting the growth of glioma and its application. Background technique [0002] Glioma, also known as glioma, or glioma for short, is derived from neuroepithelium or supporting cells. Glioma originates from glial cells in the brain and is the most common primary brain tumor in adults, accounting for 40-50% of brain tumors. Brain glial cells are composed of astrocytoma, ependymal and oligodendrocytes. Among gliomas, astrocytomas originating from astrocytes or astrocyte progenitors have the highest incidence. According to World Health Organization criteria and patient survival, astrocytomas are divided into four grades: fibrous astrocytoma (grade I), diffuse astrocytoma (grade II), anaplastic astrocytoma Cytoma (grade III), glioblastoma multiforme (GBM, grade IV). Among them, anaplastic astrocytoma and glioblastoma multiforme are malignant gliomas with poor prognosis. Especially for glioblastoma multifor...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K48/00A61P35/00C12N15/113C12N15/85C12N5/10
Inventor 黄来强陈红波申晓萌周兰贞郑义梅林王丽君
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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