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Nucleic acid molecule SEPTIN1SR2 and application in preparing anti-leukemia medicament

A nucleic acid molecule and nucleotide technology, which is applied to the nucleotide molecule and its application in the preparation of drugs for treating leukemia, can solve the problems of low bone marrow probability, low immunity, huge manpower and material resources, etc.

Inactive Publication Date: 2008-08-13
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since most of the current leukemia treatments require bone marrow transplantation, this hinders further improvement of the cure rate or remission rate
First of all, the probability of finding a matching bone marrow is not high; second, during the bone marrow transplant operation, due to the one-time killing of mature immune cells, the patient's immunity is extremely low, and it is necessary to live in a sterile room for treatment, which brings great benefits to the patient. There was a lot of pain; again, the bone marrow transplant operation consumed a lot of manpower and material resources, and brought a lot of burden and pressure to the patients and their families

Method used

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  • Nucleic acid molecule SEPTIN1SR2 and application in preparing anti-leukemia medicament
  • Nucleic acid molecule SEPTIN1SR2 and application in preparing anti-leukemia medicament
  • Nucleic acid molecule SEPTIN1SR2 and application in preparing anti-leukemia medicament

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] The expression situation in the organization of embodiment 1 Septin1

[0057] 1.1 Northern probe preparation

[0058] Clontech Multiple Tissue northen membrane was used to detect the distribution of septin1 gene in adult tissues, and the probe used was the cloned full-length open reading frame of septin1 (ie, positions 188 to 1291 in the nucleotide sequence shown in NCBI accession number NM_052838).

[0059] 1) Using specific primers of septin1 gene to amplify the PCR product from the constructed and sequenced plasmid. After the PCR product was purified with the QIAquick PCR purification kit, the concentration of the purified DNA was measured with a GeneQuant II UV spectrophotometer.

[0060] 2) After the DNA template was heat-denatured at 100°C for 2 minutes, [γ-32P]dCTP was incorporated with a random primer labeling kit using a random primer labeling method. After the labeling reaction, EDTA was added to a final concentration of 20mmol / L to stop the reaction, heat d...

Embodiment 2

[0068] Example 2 western-blot analysis of septin1 protein expression in 17 human cell lines

[0069] 1. Culture of 17 human cell lines

[0070] The selected 17 kinds of cells are all from the Cell Bank of the Chinese Academy of Sciences, and their tissue sources and culture conditions are as follows:

[0071] name

species

source of organization

Culture conditions

HEK293T

Humanity

embryonic kidney

90% DMEM high glucose complete medium, 10% fetal

Bovine serum, 37°C, 5% CO 2

HELA

Humanity

cervix

Same as HEK293T

U2OS

Humanity

muscle

90% 5A medium, 10% fetal bovine serum, 37

°C, 5% CO 2

PANC

Humanity

pancreas

Same as HEK293T

SMMC772

1

Humanity

the liver

Same as HEK293T

MCF7

Humanity

breast

Same as HEK293T

H1299

Humanity

lung

90% RPMI1640 medium, 10% fetal bovine...

Embodiment 3

[0082] Example 3 Localization of Septin1 in Jurkat cells

[0083] In the present invention, firstly, the cDNA of full-length septin1 is constructed into the Ds-Red-Cl fluorescent expression vector. The resulting RFP-septin1 recombinant was transfected into Jurkat cells, cultured for 48 hours, fixed, and observed under a confocal laser microscope. Specific steps are as follows:

[0084] (1) Collect the Jurkat cells cultured in suspension, disperse them fully, drop them on slides coated with poly-lysine, let them stand for 5 minutes, and then fix them in 4% paraformaldehyde at room temperature for 10 minutes (minutes).

[0085] (2) Wash three times with PBS, and punch holes in 0.2% Triton X-100 at room temperature for 15 minutes.

[0086] (3) Wash three times with PBS, block in TBS containing 10% horse serum and 1% BSA at room temperature for 1 hr (hour).

[0087] (4) Wash once with TTBS, add primary antibody (anti-septin1 antibody, 1:20) diluted in proportion with antibody d...

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Abstract

The invention belongs to abc warfare medicine field and relates to a nucleotide molecule and its use for preparing leukaemia medicaments. Leukaemia is a malignant tumo(u)r of hemapoietic system. At present, leukaemia can be treated by bone marrow grafting. Leukaemia brings prodigious burden and drang to patient and their family numbers. The nucleotide sequence of the invention comprises AUUCAUCGAGGAGCAAUUU, ATTCATCGAGGAGCAATTT, AAATTGCTCCTCGATGAAT or AAAUUGCUCCUCGAUGAAU. The nucleotide can halt lymphocyte abruption so the pharma-ceutical product composition comprising the nucleotide and transporting species or excipient vehicle in pharmacy can be used to be new pharma-ceutical product for treating leukaemia. The invention provides a new path and means for releasing and treating lymphocyte leukaemia.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a nucleotide molecule and its application in the preparation of medicines for treating leukemia. Background technique [0002] Leukemia is a malignant tumor of the hematopoietic system. It is characterized by decreased production of normal blood cells, qualitative and quantitative abnormalities of peripheral blood leukocytes, progressive and uncontrolled abnormal proliferation of leukocytes and their immature cells (ie, leukemia cells) in bone marrow or other hematopoietic tissues, infiltrating and damaging various tissues, produce different symptoms. According to the course of leukemia and cell morphology, leukemia can be divided into: (1) acute leukemia develops rapidly, and the bone marrow and peripheral blood are mainly abnormal primitive and immature cells. The natural course of the disease is mostly within 6 months. Acute leukemia is divided into: A, acute lymphoblastic leukemia ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C07H21/02A61K31/7088A61K48/00A61P35/02C12N15/113
Inventor 余龙余文博唐丽莎丁向明
Owner FUDAN UNIV