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Application of iMAC1 in preparation of product for treating cancer

A cancer, product technology, applied in the field of biomedicine

Active Publication Date: 2020-10-23
军事科学院军事医学研究院生物医学分析中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, autophagy plays a very important role in the course of human cancer, and the application of regulating autophagy in tumor treatment still needs more perfect research

Method used

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  • Application of iMAC1 in preparation of product for treating cancer
  • Application of iMAC1 in preparation of product for treating cancer
  • Application of iMAC1 in preparation of product for treating cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1, iMAC1 increases intracellular LC3 punctate aggregation

[0040] LC3 protein is a classic marker protein for evaluating autophagy and an important marker on the bilayer membrane of autophagosomes. If the punctate aggregation (LC3 puncta) of intracellular LC3 protein increases, the autophagy level of the cell increases.

[0041] 1. Take a 24-well plate, put a round cover glass in each well, and then inoculate GFP-LC3-HeLa cells for adherent culture.

[0042] 2. After completing step 1, add iMAC1 to each well to obtain system 1; in system 1, the concentration of iMAC1 is 0 μM, 2.5 μM or 5 μM.

[0043] 3. After completing step 2, take system 1 and place it at 37°C, 5% CO 2 Incubate for 2 h in a cell culture incubator.

[0044] 4. Take system 1 after step 3, discard the liquid phase, and add iMAC1-containing EBSS (Earle's Balanced Salt Solution) solution (as a positive control) or iMAC1-containing DMEM medium to obtain system 2. In the EBSS solution containing i...

Embodiment 2

[0055] Example 2, iMAC1 promotes the accumulation of autophagosomes in a dose- and time-dependent manner

[0056] LC3-II is the esterified form of LC3-I, which can anchor on the bilayer membrane of autophagosomes and serve as an important marker of autophagosomes.

[0057] 1. In order to study the relationship between the level of autophagy and the dose of iMAC1, the inventors of the present invention used Western blotting to detect the changes in the abundance of LC3-II protein. Specific steps are as follows:

[0058] (1) Add iMAC1 to adherent cultured HeLa cells to obtain system 1; in system 1, the concentration of iMAC1 is 0 μM, 0.5 μM, 1.0 μM, 2.5 μM, 5.0 μM or 8.0 μM.

[0059] (2) After completing step (1), take system 1 and place it at 37°C, 5% CO 2 cultured in a cell culture incubator for 24 h.

[0060] (3) After completing step (2), extract the total protein of each cell, and then use the antibody of LC3 protein or GAPDH antibody (control) as the primary antibody fo...

Embodiment 3

[0068] Example 3, iMAC1 inhibits the downstream of the autophagic flow of cells

[0069] 1. Western blotting

[0070] (1) Prepare system 1-system 4.

[0071] Take adherent cultured HeLa cells and add Baf-A1 and iMAC1 to obtain system 1. In system 1, the concentration of Baf-A1 was 100 nM, and the concentration of iMAC1 was 5.0 μM.

[0072] Take adherent cultured HeLa cells and add Baf-A1 and DMSO to obtain system 2. In system 2, the concentration of Baf-A1 was 100 nM.

[0073] Take adherent cultured HeLa cells and add DMSO and iMAC1 to obtain system 3. In system 3, the concentration of iMAC1 was 5.0 μM.

[0074] Take adherent cultured HeLa cells and add DMSO to obtain system 4.

[0075] (2) After completing step (1), take system 1-system 4 respectively and place them at 37°C and 5% CO 2 cultured in a cell culture incubator for 24 h.

[0076] (3) After completing step (2), extract the total protein of each cell, and then use the antibody of LC3 protein or Tubulin antibo...

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Abstract

The invention discloses an application of iMAC1 in preparation of a product for treating cancer. Experimental results show that the iMAC1 has the following functions: the LC3 dot aggregation in cellsis increased, the accumulation of autophagosomes is promoted and presents dose and time dependence, the autophagy flow downstream of the cells is inhibited, the average number of the autophagosomes and autophagy lysosomes of HeLa cells is significantly increased, the degradation of damaged mitochondria is inhibited, and the activity of tumor cells is significantly reduced. Therefore, the iMAC1 canbe used for preparing anti-tumor drugs and has important application value in the aspect of cancer treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of iMAC1 in preparing products for treating cancer. Background technique [0002] Inhibitor of mitochondrial apoptosis-induced channel inhibitor 1 (Inhibitor of mitochondrial apoptosis-induced channel 1, iMAC1), also known as BAX Channel Blocker, its molecular formula is C 19 h 23 Br 2 Cl 2 N 3 O, structural formula such as figure 1 Shown, the full name is 3,6-dibromoindole-alpha-(1-piperazinemethyl)-9H-carbazole-9-ethanol (1-(3,6-dibromocarbazol-9-yl)-3- piperazin-1-ylpropan-2-ol). iMAC1 is a 3,6-dibromocarbazole piperazine derivative that acts on BAX protein and can inhibit the release of cytochrome c from mitochondria. Studies have found that iMAC1 also inhibits the activity of BAK protein, and its function is mainly in two aspects: one is to bind and inhibit the activation and allosteric modification of BAX protein, and inhibit the positioning of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/496A61P35/00
CPCA61K31/496A61P35/00
Inventor 张学敏潘欣李爱玲周涛赵菲陈佳意
Owner 军事科学院军事医学研究院生物医学分析中心
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