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Application of cucurbitacin B in preparation of ferroptosis inducer and anti-nasopharyngeal carcinoma drug

A nasopharyngeal cancer and cucurbitacin technology, applied in the field of medicine, can solve the problems of small destructive power of the immune system, small bone marrow suppression, low liver damage, etc., and achieve good anti-nasopharyngeal cancer tumor effect

Inactive Publication Date: 2020-09-15
THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cucurbitacin B (Cucurbitacin B, CuB) is one of the most abundant cucurbitacin derivatives. It has been reported that cucurbitacin B can be used in the treatment of various tumors. Compared with conventional chemotherapy drugs, it has less inhibitory effect on bone marrow. , low liver damage, and low destructive power of the immune system. It has important potential as an anti-tumor drug, but there is no relevant research on its treatment of nasopharyngeal carcinoma, and its main mechanism of action is even less clear

Method used

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  • Application of cucurbitacin B in preparation of ferroptosis inducer and anti-nasopharyngeal carcinoma drug
  • Application of cucurbitacin B in preparation of ferroptosis inducer and anti-nasopharyngeal carcinoma drug
  • Application of cucurbitacin B in preparation of ferroptosis inducer and anti-nasopharyngeal carcinoma drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Cucurbitacin B inhibits proliferation of nasopharyngeal carcinoma cells

[0042] 1. Experimental materials and instruments

[0043] (1) 1640 medium, 0.25% trypsin, Gibco fetal bovine serum; 96-well plate (Corning, USA); multifunctional microplate reader (Synergy H1, BioTek, USA); biological safety cabinet (Thermo Fisher Scientific, USA) , CO2 cell incubator (Thermo Fisher Scientific, USA).

[0044] (2) Cell line: CNE1 (nasopharyngeal carcinoma cells), cultured in 1640 complete medium normally containing 10% fetal bovine serum.

[0045] (3) Solution preparation: MTT reagent, weigh 150mg MTT and dissolve it in 150mL PBS to prepare a solution with a concentration of 5mg / mL, then filter through a 0.22μm filter membrane and store in a dark place at -20°C.

[0046] 2. Proliferation inhibition test method

[0047] Collect logarithmic phase cells, inoculate 96-well cell culture plate with 5000 cells per well, fill the edge wells of the plate with sterile PBS, 5% CO...

Embodiment 2

[0050] Example 2: Cucurbitacin B induces the morphology of nasopharyngeal carcinoma cell CNE1 to exhibit ferroptosis characteristics

[0051] 1. Electron microscope observation of cell morphology

[0052] Collect logarithmic phase cells, inoculate 1 million cells in 60 mm and incubate for 24 hours. Cucurbitacin B (50 nM) was added, and culture was continued for 24 hours. Cells were collected, fixed with 2.5% glutaraldehyde for 2 hours, then fixed with 1% osmic acid for 1.5 hours, dehydrated, soaked, embedded in resin, and scanned by electron microscope for cell substructure. At the same time, the known ferroptosis inducer Erastin (10 μM) was used as a control, and the incubation time was also 24 hours.

[0053] 2. Experimental results

[0054] Electron microscope observation results such as figure 2 As shown in the electron microscope, under the action of cucurbitacin B, the mitochondria in CNE1 cells were rounded, the mitochondrial membrane shrunk, and the mitochondrial ...

Embodiment 3

[0055] Example 3: Effect of cucurbitacin B on iron content in nasopharyngeal carcinoma cells CNE1

[0056] The inventors have found in previous studies that the use of ferroptosis inhibitor deferoxamine (Deferoxamine, DFO) can effectively prevent cell death caused by cucurbitacin B. Therefore, we speculate that the cell death induced by cucurbitacin B is closely related to ferroptosis. In order to further verify that cucurbitacin B has the effect of inducing ferroptosis, the intracellular iron ion content was detected after cucurbitacin B was treated with drugs.

[0057] 1. Detection of intracellular iron ion content

[0058] The logarithmic phase cells were collected and seeded in 6-well cell culture plates at 300,000 cells per well and incubated for 24 hours. 1-50nM cucurbitacin B was added to incubate the cells for 24 hours, the cells were homogenized and lysed, and the iron ion content in the cells was detected according to the instructions of the iron ion detection kit (...

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Abstract

The invention discloses an application of cucurbitacin B in preparation of a ferroptosis inducer and an anti-nasopharyngeal carcinoma drug. The research shows that cucurbitacin B can induce ferroptosis for the first time; death of nasopharyngeal carcinoma cells is regulated and controlled in a novel cell death promoting mode; and further research finds that cucurbitacin B can promote accumulationof iron ions, reduce the content of glutathione in the cells and down-regulate expression of GPX4 in nasopharyngeal carcinoma cells CNE1, so that lipid peroxide is increased, and iron death of the cells is induced. In in-vitro and in-vivo studies, cucurbitacin B induces the CNE1 cell cycle to be blocked in the G2 / M period and inhibits cell migration and invasion. In addition, cucurbitacin B can significantly inhibit growth of mouse nasopharyngeal carcinoma tumors, and has no obvious toxic or side effect. The invention provides a new choice, namely a ferroptosis inducer, for a medicine for treating nasopharyngeal carcinoma; and meanwhile, a natural novel medicine source, namely cucurbitacin B, is provided for a ferroptosis inducer and treatment of nasopharyngeal carcinoma. The cucurbitacinB is efficient and safe and is free of toxic and side effects.

Description

technical field [0001] The invention belongs to the technical field of medicine. More specifically, it relates to the application of cucurbitacin B in the preparation of ferroptosis inducers and anti-nasopharyngeal cancer drugs. Background technique [0002] Nasopharyngeal carcinoma is a malignant tumor that occurs on the top and side walls of the nasopharyngeal cavity. Nasopharyngeal carcinoma mainly occurs in the five southern provinces of my country, namely Guangdong, Guangxi, Hunan, Fujian and Jiangxi, accounting for the first place in local head and neck malignant tumors, and Southeast Asian countries are also high-incidence areas. At present, there is no particularly effective drug for the treatment of nasopharyngeal carcinoma. Radiation therapy is the first choice for the treatment of nasopharyngeal carcinoma. However, for patients with highly differentiated carcinoma, late course of disease and recurrence after radiotherapy, surgical resection and chemotherapy are s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/575A61P35/00A61P11/02A61P11/04
CPCA61K31/575A61P11/02A61P11/04A61P35/00
Inventor 梁宝霞严君廖升荣
Owner THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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