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Use of CMTM1-V5 gene and its encoded protein

A 1.cmtm1-v5, coding technology, applied in the field of leukemia treatment, can solve the problem of poor efficacy of lymphoma

Inactive Publication Date: 2016-08-24
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bortezomib is a protease inhibitor that has been used clinically in recent years. It mainly blocks the activation of NF-κB by preventing the degradation of I-κB, thereby inhibiting cell proliferation, causing cell apoptosis, and playing an anti-tumor effect. It can make tumor cells more sensitive to chemotherapy, radiation therapy and immunotherapy, and overcome the resistance of cells to drugs. It has shown good efficacy and clinical tolerance in the treatment of myeloma and some lymphoma subtypes, but it is not effective for How much lymphoma does not respond well

Method used

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  • Use of CMTM1-V5 gene and its encoded protein
  • Use of CMTM1-V5 gene and its encoded protein
  • Use of CMTM1-V5 gene and its encoded protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1. Overexpression of CMTM1-v5 can inhibit the growth of Jurkat and Raji cells

[0085] The proliferation of Jurkat and Raji cells overexpressing CMTM1-v5 was measured by MTT method and cell viability counter.

[0086] MTT is a light yellow methyl tetrazolium salt. The mitochondrial dehydrogenase of living cells can produce blue crystalline formazan with MTT. The formazan can be dissolved in dimethyl sulfoxide (DMSO) for color comparison. The amount of formazan production is directly proportional to the metabolism of living cells, and only living cells have this reaction. The result is measured with a microplate reader, which can reflect the strength of cell metabolic activity according to the depth of color. Inoculate the transfected cells in a 96-well plate at a density of 2000 cells / well. Each group of cells has 5 parallel re-wells and cultured continuously for 6 days. Add MTT every 24 hours and incubate at 37°C, 5% CO2. 4 -6 hours later, add the cell lysate, mea...

Embodiment 2

[0089] Example 2: Overexpression of CMTM1-v5 can induce apoptosis of Jurkat and Raji cells

[0090] Phosphatidylserine (PS) eversion from the cytoplasmic side of the cell membrane to the outer surface of the cell membrane is one of the typical biochemical characteristics of apoptosis. Annexin-V can specifically bind to the PS on the outer side of the membrane. The fluorescein-labeled Annexin-V combined with flow cytometry can be used to quantitatively determine cell PS eversion, and use propidium iodide (PI) staining to determine the cell membrane Completeness. Such as( figure 2 ), the abscissa represents the fluorescence intensity of FITC-Annexin-V, the ordinate represents the fluorescence intensity of PI, the lower left area represents normal cells, the lower right, upper right, and upper left areas represent early apoptosis, mid-late apoptosis, and death, respectively Cell. Jurkat cells were transiently transfected with pcDB, pcDB-CMTM1-v5 and Bax plasmids. About 8 hours af...

Embodiment 3

[0091] Example 3. Overexpression of CMTM1-v5 induces a decrease in mitochondrial transmembrane potential (Δψm)

[0092] Mitochondria are one of the key regulators of apoptosis signaling pathways. The impaired mitochondrial transmembrane potential (Δψm) is one of the earliest biochemical characteristics of mitochondrial-mediated apoptosis. In order to determine whether CMTM1-v5 mediated apoptosis is involved in mitochondria, we used DIOC6(3) dye on Jurkat cells transfected with CMTM1-v5, and analyzed the change of Δψm with flow cytometry. The cells were harvested in a centrifuge tube at different time points after transfection, centrifuged at 1500 rpm for 5 minutes, and the supernatant was discarded; the cells were washed twice with PBS and resuspended in 400l PBS to prepare a single cell suspension. After adding 20nM DiOC6(3) and incubating at 37°C for 15 minutes in the dark, flow cytometry analysis was performed immediately. Harvest 1×104 cells and calculate the percentage of c...

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Abstract

The invention relates to a use of a CMTM1-V5 polypeptide or a CMTM1-V5 polypeptide encoding polynucleotide as a biological medicine in treatment on lymphocytic leukemia and other leukemia. The invention also relates to a use of the CMTM1-V5 polypeptide or the CMTM1-V5 polypeptide encoding polynucleotide in treatment on inflammation and autoimmune diseases through NFKB and NFAT activation inhibition. The invention also relates to a pharmaceutical composition for treating leukemia, inflammation and autoimmune diseases.

Description

technical field [0001] The present invention relates to a new application of CMTM1-v5, specifically, the present invention relates to the preparation of CMTM1-v5 polypeptide or polynucleotide encoding it for the treatment of leukemia, inflammation and autoimmune diseases. Background technique [0002] Lymphoma is a malignant tumor of the blood lymphatic system. In recent years, with the aging of the society and the deterioration of the environment, the incidence of lymphoma has increased year by year. According to the statistics of new cancer cases in the United States in 2012, non-Hodgkin's lymphoma The annual incidence rate of hematological malignancies exceeds the sum of all hematological malignancies, ranking seventh among all tumors. The incidence of malignant lymphoma in my country is also increasing year by year, and it has become an important disease that threatens national health. There are many subtypes of lymphoma with strong heterogeneity, and some patients are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/17A61P35/02
Inventor 王露马大龙张国英
Owner PEKING UNIV
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