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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
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 ...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com