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Apoptosis-associated gene and medicaments for treating leucocythemia using the gene as target

A leukemia and gene technology, applied in gene therapy, drug combination, genetic engineering, etc., can solve the problem of poor leukemia effect, achieve the effect of blocking proliferation, increasing apoptosis rate, and ensuring accurate inheritance

Inactive Publication Date: 2008-12-31
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide an apoptosis-related gene and a drug targeting it for treating leukemia. The drug for treating leukemia should solve the problem of poor efficacy of drugs and technical means in the prior art for treating leukemia. technical issues

Method used

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  • Apoptosis-associated gene and medicaments for treating leucocythemia using the gene as target
  • Apoptosis-associated gene and medicaments for treating leucocythemia using the gene as target
  • Apoptosis-associated gene and medicaments for treating leucocythemia using the gene as target

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Obtaining the full-length sequence of the apoptosis-related gene PNAS-2 and its gene informatics research

[0036] 1. Cell culture: NB4 acute promyelocytic leukemia cell line, provided by Shanghai Institute of Hematology. Cells were placed in RPMI 1640 containing 10% calf serum, cultured at 37°C in 5% CO2 humidified air, passaged once every 3 days, and cells in logarithmic growth phase were used for experiments.

[0037] 2. Total RNA extraction, RNA integrity detection: take 5×10 6 Add 1 NB4 cells into a 15ml centrifuge tube, centrifuge at 1500rpm in a cryogenic centrifuge at 4°C, and discard the supernatant. The cell pellet was washed with ice-cold PBS buffer (pH 7.4), centrifuged again, and the supernatant was discarded. Add 1ml Trizol, pipette and mix well, then transfer to a 1.5ml EP tube, let stand at room temperature for 5 minutes, then add 200μl of chloroform, shake vigorously for 15-30 seconds to fully mix the liquid, and let stand at room temperatur...

Embodiment 2

[0064] Example 2 Research on the relationship between the apoptosis-related gene PNAS-2 of the present invention and cell apoptosis

[0065] Although the original PNAS-2 was named "apoptosis-related gene", there is no experimental study on the relationship between this gene and apoptosis. Although we found that the full-length PNAS-2 and CHMP5 are homologous genes, there is no experimental study on the relationship between CHMP5 and apoptosis.

[0066] 1. The relationship between the expression of PNAS-2 and apoptosis

[0067] 1.1 Recovery and purification of high-fidelity PCR and PCR products: According to the full-length sequence of PNAS-2 obtained by our previous 5′RACE, primers were designed using Primer Premier 5, and BstXI enzyme was added to the 5′ ends of the upstream and downstream primers Cutting site and protective base (lowercase part below), the upstream primer is: 5′tag cca gtg tgg tgg TGC TCAAGATGAACC GAC TCT 3′, the downstream primer is: 5′tgt cca ctg tgc tgg ...

Embodiment 3

[0083] Example 3 The relationship between the expression of the apoptosis-related gene PNAS-2 of the present invention and the onset of leukemia

[0084] At present, there is no experimental study on the relationship between PNAS-2 and its homologous genes such as CHMP5 and the pathogenesis of leukemia.

[0085] 1.RealTime PCR method to detect the expression of PNAS-2 in patients

[0086] 1.1 Case collection: All patients were inpatients and outpatients of Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine. These included: ①. 44 cases of first-episode acute leukemia patients, including 9 cases of acute lymphoblastic leukemia; 35 cases of acute myeloid leukemia, including 3 cases of M2 patients, 8 cases of M3 patients, 18 cases of M4 patients, and 6 cases of M5 patients. ②, 27 cases of complete remission (CR) samples, including 4 cases of M2 patients, 18 cases of M3, 1 case of M4, 2 cases of M5 and 2 cases of ALL. ③. There were 12 relapse samples, in...

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Abstract

The invention provides an apoptosis-related gene, the gene contains a base sequence shown in SEQ ID NO: 1 or SEQ ID NO: 2. The invention also provides a carrier containing at least one target sequence of RNA interference for inhibiting the apoptosis-related gene. The invention also provides a cell which contains the carrier. The invention also provides a drug for curing leukemia, and the drug contains an effective amount of the RNA interference target sequence, or the carrier (containing the RNA interference target sequence) or the cell. The gene applies the RNA interference technique to inhibit PNAS-2, and has obvious effect to the treatment of leukemia.

Description

Technical field: [0001] The invention relates to the field of bioengineering, in particular to gene recombination technology, in particular to an apoptosis-related gene and a drug for treating leukemia with the target. Background technique: [0002] Apoptosis and cell proliferation are the basic phenomena of life and the basic measures to maintain the dynamic balance of the number of cells in the body. Eliminate aging and diseased cells through apoptosis to ensure the health of the body. Like cell proliferation, apoptosis is also a precise process regulated by genes. At present, it is recognized that inhibition of cell apoptosis is one of the important reasons for the occurrence of malignant tumors such as leukemia. The main means of clinical treatment of malignant tumors, chemotherapy, is to induce apoptosis of tumor cells to achieve the purpose of treatment. [0003] Leukemia is a malignant tumor of the hematopoietic system, which is caused by excessive proliferation and ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07H21/02A61K31/7088A61K48/00A61P35/02
Inventor 王海嵘陈芳源
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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