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Application of satb1 in the treatment of cutaneous T-cell lymphoma

A lymphoma, cell technology, applied in gene therapy, medical preparations containing active ingredients, antitumor drugs, etc., can solve problems such as unclear role of SATB1

Active Publication Date: 2016-02-03
PEKING UNIV FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The role of SATB1 in peripheral mature T cells and T-cell tumors remains unclear

Method used

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  • Application of satb1 in the treatment of cutaneous T-cell lymphoma
  • Application of satb1 in the treatment of cutaneous T-cell lymphoma
  • Application of satb1 in the treatment of cutaneous T-cell lymphoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1. SATB1 is specifically highly expressed in CD30+ neoplastic T cells of cutaneous T-cell lymphoma

[0075] 1. Immunohistochemical staining

[0076] Immunohistochemical staining was performed on paraffin specimens of MF large cell transformation (MF-LCT), lymphomatoid papulosis (LyP), and primary cutaneous anaplastic large cell lymphoma (PCALCL), as follows:

[0077] (1) Embed the specimen in paraffin, make 5 μm slices, routinely dewax into water, put in xylene I, II for 10 minutes each, absolute ethanol I, II, ethanol aqueous solution I, 95% by volume II, 80% ethanol water solution by volume for 5 minutes each, fully hydrated with distilled water for 3-5 minutes, and rinsed with PBS (pH7.4) 3 times, 5 minutes each time.

[0078] (2) Antigen retrieval: Put the tissue slices into 0.01mol sodium citrate buffer, heat in a microwave oven on high heat until boiling, then switch to medium heat and heat for 10 minutes, take it out and let it stand to cool to room temp...

Embodiment 2

[0141]Example 2. The expression of SATB1 increases with the progression of cutaneous T-cell lymphoma (CTCL)

[0142] The sources and characteristics of the following cell lines are shown in Table 2.

[0143] Table 2 The source and characteristics of the cell lines used

[0144]

[0145] The cell culture used in the embodiment adopts RPMI1640 liquid culture medium, adds the fetal calf serum that volume percentage is 10%, 100U / ml penicillin, 0.1mg / ml streptomycin, and the cell is the suspension type growth in the culture medium, inserts Constant temperature 37°C, humidified 5% CO 2 Cell incubator, according to the state of the cells, replace the medium for passage.

[0146] 1. Extract the total RNA of Hut78, Jurkat, Mac-1, Mac-2A and Mac-2B and synthesize the first strand of cDNA.

[0147] 2. Real-time fluorescent quantitative PCR

[0148] Using the cDNA obtained in step 1 as a template, PCR amplification was performed with specific primers for the SATB1 gene, and the pro...

Embodiment 3

[0195] Example 3. The high expression of SATB1 promotes the malignant biological phenotype of cutaneous T-cell lymphoma disease

[0196] 1. Lentiviral vector

[0197] The GV118 lentiviral vector is a cloning vector, and its structure is as follows: Figure 6 shown.

[0198] Figure 6 Among them, the full length of the GV118 vector is 10307bp, and HpaI and XhoI are multiple cloning sites. GFP was used as a fluorescent marker for the selection of transfected cells. The ampicillin resistance gene can be used as a screening gene for cell clones.

[0199] 2. Construction of RNA interference lentiviral vector

[0200] (1) Design of shRNA sequence:

[0201] For the SATB1 gene sequence, use the RNA interference sequence design principles provided in the public website to design the following RNA (shRNA) sequence interference target sequence:

[0202] shRNA0:

[0203] 5'-GCGTTAACCGATTCGGTGCTACTCTATGCAAGAGATCGAGTAGCACCGAATCGCTCGAG-3'; (SEQ ID No. 7)

[0204] shRNA1:

[0205] 5'-...

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Abstract

The invention discloses application of SATB1 to treatment of cutaneous T cell lymphomata. The invention discloses application of a product taking the following any one substance as a target to preparation of medicaments for preventing and / or treating cutaneous T cell lymphomata: (1) a protein shown in SEQ ID No.1; (2) a DNA molecule shown in SEQ ID No.2; (3) a DNA molecule shown in nucleotides from 1736th to 4024th beginning from the 5'-end in SEQ ID No.2. SATB1 protein is specifically expressed in CD30+ tumorous T cells in cutaneous T cell lymphomata, the expression up-regulation of the SATB1 protein plays a major role in attack and development of the cutaneous T cell lymphomata through promoting the cell cycle development, and SATB1 serving as a target has a bright application prospect in treatment of cutaneous T cell lymphomata.

Description

technical field [0001] The invention relates to the application of a protein and its gene as a target in the treatment of skin T-cell lymphoma; in particular, it relates to the application of SATB1 in the treatment of skin T-cell lymphoma, which belongs to the field of biotechnology. Background technique [0002] Cutaneous T-cell lymphoma (CTCL) is a type of extra-nodal Non-Hodgkin's lymphoma (NHL), which is a T lymphocyte that originates in the skin and homing from the skin The diseases caused by clonal hyperplasia consist of a group of diseases with different clinical manifestations, disease course and prognosis, and treatment methods. Among them, Mycosis Fungoides (MF) and primary skin CD30+ T lymphocyte proliferative disease (CD30+ cutaneous lymphoproliferative disease, CD30+ CLPD) are the most common types, and the incidence of both accounts for more than 80% of CTCL. CD30+CLPD is a disease caused by the clonal proliferation of CD30-expressing T lymphocytes in the skin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K39/395A61K48/00A61P35/00
Inventor 汪旸涂平张高磊
Owner PEKING UNIV FIRST HOSPITAL
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