Antitumor drug and acting target thereof

An anti-tumor drug and target technology, applied in the field of biomedicine, can solve problems such as large side effects, inability to prevent tumor recurrence and distant metastasis, and short development history of chemotherapy

Active Publication Date: 2018-12-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Tumor is a major disease that endangers human health and life. At present, tumor treatment is mainly based on surgical treatment, adjuvant chemotherapy, radiotherapy or biological treatment. It has been proved by clinical practice that some early malignant tumors or certain specific tumors can be cured by certain methods. However, surgical treatment alone cannot prevent tumor recurrence and distant metastasis. Although radiotherapy can cure a variety of tumors, it still has certain limitations. As a systemic treatment method, it can kill the tumor cells in the patient's body to the greatest extent compared with other methods, and chemotherapy is transitioning from palliative treatment to radical treatment, so chemotherapy plays a very important role in tumor treatment. status
However, the selective inhibitory effect of chemotherapy on tumor cells is poor, and systemic drug side effects are large, including gastrointestinal reactions, bone marrow suppression, hair loss, adverse reactions of vital organs such as heart, liver and kidney, and long-term carcinogenicity and teratogenicity, which limit its application. Therefore, improving the efficacy of chemotherapy drugs is of great significance for tumor treatment

Method used

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  • Antitumor drug and acting target thereof
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  • Antitumor drug and acting target thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Using female SPF grade C57BL / 6 as the experimental object, the mouse colon cancer cell line MC38 was used to construct a tumor-bearing mouse model, and the enzyme-linked immunosorbent assay double antibody sandwich method was used to detect CD19 in serum and spleen tissue supernatant + EVs content.

[0037] CD19-positive exosomes in serum were detected by enzyme-linked immunosorbent assay (ELISA) + EVs) content. Specific steps are as follows:

[0038] (1) Collect mouse spleen tissue and squeeze it with the plunger of a syringe to make a tissue suspension; (2) Transfer to a 15ml conical test tube and allow the large pieces to settle to the bottom of the test tube or filter through a nylon filter to obtain a single Cell suspension; (3) Centrifuge the cell suspension at 1500rpm at 4°C for 4-5 minutes, and collect the supernatant; (4) Take the peripheral blood of the mouse and put it in a 1.5ml tube, let it stand at room temperature for 2 hours, overnight at 4°C, and coll...

Embodiment 2

[0043] Spleen B cells of normal mice and tumor-bearing mice were counted and analyzed by flow cytometry. Proceed as follows:

[0044] (1) Collect mouse spleen tissue and squeeze it with the plunger of a syringe to make a tissue suspension; (2) Transfer to a 15ml conical test tube and allow the large pieces to settle to the bottom of the test tube or filter through a nylon filter to obtain a single Cell suspension; (3) Centrifuge the cell suspension at 1500rpm at 4°C for 4-5 minutes, discard the supernatant; (4) For the precipitate obtained in the previous process, lyse the red blood cells with red blood cell lysate, add twice the volume of serum-free 1640 medium, centrifuge under the same conditions, discard the supernatant; (5) resuspend the sample with 2ml PBS buffer, and perform cell counting; (6) centrifuge the cells again, discard the supernatant, and resuspend the cells so that the number of cells reaches 1×10 7 pcs / ml; (7) take out 1×10 7 The cells were centrifuged t...

Embodiment 3

[0047] Since the Rab family has a regulatory effect on the secretion of EVs, real-time fluorescent quantitative PCR detection was performed on the Rab family-related factors in B cells to screen the factors that affect the secretion of vesicles. The specific method is as follows:

[0048] (1) Take the spleen tissues of tumor-bearing mice and normal mice, and use the method in Example 2 to obtain spleen single cell suspension; Methods: Sorting mouse B cells; (3) Dissolve the sorted cells in 500 μl Trizol reagent (Takara, Cat. No. 9109), overnight at 4°C, and extract RNA the next day; (4) Add 100 μl chloroform to each tube, vortex Rotate and oscillate for 1 minute; (5) shake vigorously to mix the reagents for 1 minute; (6) stand at room temperature for 10 minutes, then centrifuge at 12000G and 4°C for 15 minutes; (7) draw the upper aqueous phase into a new EP tube, add Equal volume of isopropanol, upside down 6 times; (8) Stand at room temperature for 10 minutes, then centrifug...

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Abstract

The invention discloses an antitumor drug and an acting target thereof. Researches discover that CD19+EVs may act as a novel clinical inspection index for in-vitro early tumors; by comparing tumor-bearing mice to normal mice, it is discovered that the level of CD19+EVs in the serum of the tumor-bearing mice has a significant rise; factors that affect the level of CD19+EVs are further studied, andit is discovered that the level of Rab27a protein in B cells of the tumor-bearing mice rises, and the B cells are stimulated to release more EVs. By using siRNA interference specific to Rab27a gene toreduce the expression level of Rab27a protein in cells, it is possible to effectively inhibit B cells from secreting CD19+EVs; the results show that chemotherapy effect can be greatly improved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an action target of an antitumor drug and the antitumor drug. Background technique [0002] Tumor is a major disease that endangers human health and life. At present, tumor treatment is mainly based on surgical treatment, adjuvant chemotherapy, radiotherapy or biological treatment. It has been proved by clinical practice that some early malignant tumors or certain specific tumors can be cured by certain methods. However, surgical treatment alone cannot prevent tumor recurrence and distant metastasis. Although radiotherapy can cure a variety of tumors, it still has certain limitations. As a systemic treatment method, it can kill the tumor cells in the patient's body to the greatest extent compared with other methods, and chemotherapy is transitioning from palliative treatment to radical treatment, so chemotherapy plays a very important role in tumor treatment. status. Howeve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/7105A61K45/06A61P35/00
CPCA61K31/7105A61K45/00A61K45/06A61P35/00
Inventor 蔡志坚王建莉张方慧杨云山李荣荣
Owner ZHEJIANG UNIV
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