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Application of aspirin in preparation of medicine for causing tumor immunogen cell death

An aspirin, immunogenic technology for use in biotechnology and medicine

Pending Publication Date: 2022-08-05
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method still has several limitations, including infiltration of tumor-infiltrating lymphocytes (TIL), expression of PD-L1, hypermutation, and expression of neoantigens

Method used

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  • Application of aspirin in preparation of medicine for causing tumor immunogen cell death
  • Application of aspirin in preparation of medicine for causing tumor immunogen cell death
  • Application of aspirin in preparation of medicine for causing tumor immunogen cell death

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Using Aspirin to inhibit the activity of CT26 cells and cause apoptosis

[0034] The apoptosis promotion and inhibition of CT26 colorectal cancer cells with different concentrations of Aspirin were determined by cell activity assay and apoptosis marker detection, including the following steps:

[0035] (1) Take the cells in the logarithmic growth phase and count them with a hemocytometer after digestion;

[0036] (2) Add 5×10 to the 96-well plate 3 100 μL of cells per well; add different concentrations of Aspirin for 24 hours after adhering for 2 to 4 hours;

[0037] (3) Add 10 μL of CCK8 reagent to each well, and use a microplate reader to detect the absorbance of each well at 450 nm after 24 hours;

[0038] (4) Draw the standard curve and calculate the inhibitory effect of different concentrations of Aspirin on tumor cell proliferation.

[0039] The inhibitory effect of Aspirin obtained according to the above method on tumor cell proliferation is as foll...

Embodiment 2

[0046] Example 2. Using Aspirin to induce immunogenic cell death of CT26 colorectal cancer cells

[0047] 1. CT26 colon cancer cells were treated with different concentrations of Aspirin to detect the expression of CRT, HSP70 and HMGB1

[0048] (1) HMGB1 level detection:

[0049] Will 5x10 5 The tumor cells were seeded in each well of a 24-well plate and treated with Aspirin for 24 hours. The cell culture medium was collected, and the HMGB1 content in the culture medium was detected by ELISA detection kit.

[0050] (2) CRT and HSP70 detection:

[0051] Will 5x10 5 The tumor cells were seeded in each well of a 24-well plate and treated with Aspirin for 24 hours. The cell culture medium was collected, washed and incubated with the corresponding antibodies, and the expression of surface molecules was detected by flow cytometry.

[0052] 2. The experimental results obtained by the above method are as follows: figure 2 As shown, we treated CT26 colon cancer cells with Aspiri...

Embodiment 3

[0053] Example 3. Using Aspirin to induce endoplasmic reticulum stress in tumor cells

[0054] 1. Detection of intracellular ATP level:

[0055] Will 5x10 5 The tumor cells were seeded in each well of a 6-well plate and treated with Aspirin for 24 hours. The cell culture medium was collected, and the ATP content in the culture medium was detected with an ATP detection kit.

[0056] The experimental results obtained by the above method are as follows: image 3 A shown. The changes of cellular ATP content after different concentrations of drug treatment were detected. The results suggest that Aspirin has a certain inhibitory effect on the ATP secretion of CT26 colon cancer ( image 3 A).

[0057] 2. Detection of the expression of endoplasmic reticulum stress-related proteins

[0058] Will 5x10 5 The tumor cells were seeded in each well of a 6-well plate and treated with Aspirin for 24 hours. Cells were harvested and lysed to collect protein for quantification. The rela...

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Abstract

The invention provides an application of aspirin in preparation of a medicine for causing tumor immunogen cell death. The application of the small-molecule inhibitor Aspirin for causing tumor immunogen cell death so as to inhibit tumor growth is found for the first time; aspirin can cause apoptosis of colorectal cancer cells and endoplasmic reticulum stress in vitro and induce PD-L1 expression up-regulation, and Aspirin has a certain inhibition effect on occurrence and development of tumors when used alone or combined with a PD-1 antibody for treatment or an antigen vaccine in vivo; by adjusting tumor metabolism and improving tumor microenvironment, inhibition on T cell functions is reduced, T cell reactivity is improved, immunotherapy efficiency is enhanced, and a new target and a new thought are provided for tumor therapy.

Description

technical field [0001] The invention relates to the fields of biotechnology and medical technology, in particular to the application of aspirin in the preparation of a drug for causing tumor immunogenic cell death. Background technique [0002] In recent years, immunotherapy against tumors has made great progress, and immune checkpoint inhibitors have been shown to play an important role in a variety of tumors. However, this approach still has several limitations, including tumor-infiltrating lymphocyte (TIL) infiltration, PD-L1 expression, hypermutation, and neoantigen expression. Immunogenic antigens can be recognized by TIL, so enhancing the immunogenicity of tumors plays an important role in the effect of immunotherapy. [0003] Immunogenic cell death (ICD) is a way of tumor cell death that can activate the body's anti-tumor response. Tumor cells with immunogenic cell death can express a variety of related molecules, including calreticulin (CRT), heat-shock protein 70 ...

Claims

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Application Information

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IPC IPC(8): A61K31/616A61P35/00A61P37/04
CPCA61K31/616A61P35/00A61P37/04
Inventor 殷雷雷骏方嘉凌
Owner WUHAN UNIV
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