Use of MDM2 inhibitor Nutlin-3a in preparing medicine for apoptosis of cancer cells induced by activating endoplasmic reticulum stress

An endoplasmic reticulum stress, MDM2 technology, applied in the application field of cancer cell apoptosis drugs, to achieve the effect of enhancing anti-tumor effect, inducing apoptosis and inhibiting growth

Active Publication Date: 2021-06-04
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many anti-tumor drugs exert their anti-tumor effects by inducing apoptosis, among which the MDM2 inhibitor Nutlin-3a induces apoptosis of colon cancer cells through the exogenous death receptor pathway is rarely reported
[0006] In addition, so far, there have been no reports at home and abroad on the role of the MDM2 inhibitor Nutlin-3a in activating endoplasmic reticulum stress to regulate the apoptosis pathway of exogenous death receptors

Method used

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  • Use of MDM2 inhibitor Nutlin-3a in preparing medicine for apoptosis of cancer cells induced by activating endoplasmic reticulum stress
  • Use of MDM2 inhibitor Nutlin-3a in preparing medicine for apoptosis of cancer cells induced by activating endoplasmic reticulum stress
  • Use of MDM2 inhibitor Nutlin-3a in preparing medicine for apoptosis of cancer cells induced by activating endoplasmic reticulum stress

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Nutlin-3a can effectively reduce cell survival rate, inhibit cell proliferation, and induce colon cancer cell apoptosis.

[0053] 1. Nutlin-3a is a high-efficiency, non-peptide MDM2-p53 small molecule inhibitor. Its molecular formula is: C30H30Cl2N4O4, its molecular weight is 581.49, and its structural formula is as follows:

[0054]

[0055] The above compounds are used as MDM2-p53 small molecule inhibitors. The commercial name of this compound is Nutlin-3a, which can be purchased from Sigma-Aldrich Biochemical Reagent Company, USA.

[0056] 2. In vitro culture of colon cancer cells

[0057] The cells used in this experiment were P53 wild-type human colon cancer cells LOVO, using DMEM containing 10% fetal bovine serum (fetal bovine serum, FBS, Gibco) and medium containing 1% penicillin-streptomycin double antibody The cells were cultured in a 37°C, 5% CO2 incubator. When the cells grew to about 80% of the bottom of the culture dish, they were digested with 0.1% tr...

Embodiment 2

[0069] Nutlin-3a induces colon cancer cell apoptosis through the extrinsic death receptor apoptosis pathway.

[0070] 1. Nutlin-3a can induce the up-regulation of expression levels of death receptor pathway-related genes

[0071] Colon cancer cells RKO, HCT116, and LOVO cells in the logarithmic growth phase were digested with trypsin, and were mixed with 4×10 5 / Cells are evenly spread in a six-well plate. After the cells are completely adhered to the wall, replace with Nutlin-3a diluted in fresh medium with a concentration of 0 μM, 35 μM, 50 μM, and 75 μM. Continue to culture in the incubator for 20 hours, collect the cells, and use Trizol Cellular RNA was extracted using the Nano Drop 2000 spectrophotometer, and the RNA concentration was measured using a Nano Drop 2000 spectrophotometer. 1ug RNA was taken according to PrimeScript TM RT Master Mix (Perfect Real Time) Reverse Transcription Kit Instructions reversed to cDNA, according to Premix Ex Taq TM II (Tli RNaseH Plus) ...

Embodiment 3

[0085] Nutlin-3a induces endoplasmic reticulum stress by increasing intracellular calcium ion concentration

[0086] 1. Nutlin-3a can induce up-regulation of endoplasmic reticulum stress-related proteins

[0087] RKO, HCT116 and LOVO cells in the logarithmic growth phase were digested with trypsin, and inoculated evenly into four 6cm small dishes at 5×106 / dish, and divided into four groups according to the concentration of Nutlin-3a: 0 μM, 35 μM, 50 μM, 75 μM; continue to culture in the incubator for 20 hours, collect cells and extract proteins, and use Western blotting to detect the expression of endoplasmic reticulum stress-related proteins XBP1α, BIP, p-EIF-2α, ATF4, and CHOP. result Figure 12 As shown, Nutlin-3a can effectively up-regulate the expression level of endoplasmic reticulum stress-related proteins in a concentration-dependent manner.

[0088] 2. Nutlin-3a can increase intracellular calcium ion concentration

[0089] Colon cancer cells in the logarithmic grow...

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Abstract

The invention discloses use of an MDM2 inhibitor Nutlin-3a in preparing a medicine for apoptosis of cancer cells induced by activating endoplasmic reticulum stress. Experiments prove that the MDM2 inhibitor Nutlin-3a can induce the occurrence of endoplasmic reticulum stress by increasing the concentration of calcium ions in cytoplasm, so as to activate a death receptor pathway to induce apoptosis of colon cancer cells. The invention further discloses an anti-tumor pharmaceutical composition of the MDM2 inhibitor Nutlin-3a and an endoplasmic reticulum stress activator. Both in-vivo and in-vitro experiments prove that the pharmaceutical composition has a remarkable synergistic effect and a good anti-tumor effect in the aspect of colon cancer treatment.

Description

technical field [0001] The present invention relates to the field of colon cancer cell endoplasmic reticulum stress and combined medicine, more specifically, relates to the application of MDM2 inhibitor Nutlin-3a in the preparation of drugs for activating endoplasmic reticulum stress-induced apoptosis of cancer cells. Background technique [0002] Colorectal cancer (CRC) is one of the most common malignant tumors of the digestive system, accounting for about 10% of the world's annual diagnosed cancer and cancer-related deaths, with more than 1 million new cases each year. The occurrence and development of colorectal cancer is a complex process involving multiple factors, such as up-regulation of PI3K / AKT, P53 gene mutation and other factors may induce and promote the occurrence of colon cancer. [0003] Mouse double minute 2 (MDM2) is the most important negative regulator of p53. Studies have shown that the loss of p53 wild-type function exists in many cancer patients. Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/496A61K31/7072A61K31/365A61P35/00A61P1/00
CPCA61K31/496A61K31/7072A61K31/365A61P35/00A61P1/00A61K2300/00
Inventor 钟加滕陆漫漫李娜贾慧婕张哲莹苏蔚赵铁锁陈志国
Owner XINXIANG MEDICAL UNIV
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