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Application of alpha-NETA in induction of cell death

A technology of cell death and induction of cells, applied in the field of cell death induction, can solve problems such as poor effect and slow death of tumor cells

Pending Publication Date: 2019-09-17
乔联桥 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the complexity of the mechanism of cell death, although many drugs are currently developed for cell death, the effect is often poor or the death rate of tumor cells is slow. Therefore, research and regulation of various cell death mechanisms and intervention methods are still needed in this field , thus helping to solve various diseases caused by cell death

Method used

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  • Application of alpha-NETA in induction of cell death
  • Application of alpha-NETA in induction of cell death
  • Application of alpha-NETA in induction of cell death

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0243] Example 1 ɑ-NETA causes ovarian cancer cell death

[0244] After fully dissolving ɑ-NETA, place it in an EP tube and store it in a 4-degree refrigerator for later use. The mother liquor concentration is 25mg / ml. SKOV3 was treated with different concentrations of α-NETA (0, 0.25ug / ml, 0.5ug / ml, 0.75ug / ml, 10ug / ml, 12.5ug / ml), and observed under an inverted microscope as shown in the figure ( figure 1 A) (magnification 10x objective lens).

[0245] Ho8910, Ho8910pm, A2780 cell lines were tested for cytotoxicity using the CCK8 method ( figure 1 C). At the same time, the IC 50 calculate( figure 1 C) (1 μmol ɑ-NETA = 369.24 mg; 1 mg ɑ-NETA = 2.7 μmol).

[0246] Use concentration 12.5ug / ml (3.38×10 -2 μmol / ml conversion value) of ɑ-NETA blank culture solution treated Ho8910 for 24 hours (the same concentration without drug vehicle as the control group), the mortality was determined by flow cytometry ( figure 1 B). The experiment was repeated three times independently...

Embodiment 2

[0248] Example 2 ɑ-NETA can cause ovarian cancer cell membrane bubbling and cytoplasmic leakage

[0249] After Ho8910 was treated with 12.5 μg / mlɑ-NETA for 24 hours, it was observed under an inverted microscope (40 times objective lens), and the negative treatment group was used as a control, as shown in the figure ( figure 2 A).

[0250] Cell death-associated blebs can be seen within the black box ( figure 2 A), the "fried egg-like" appearance associated with the late stage of pyroptosis was simultaneously observed under a fluorescent microscope ( figure 2 B).

[0251] It was observed more finely with a transmission electron microscope, and normal ovarian cancer cells were used as a control as shown in the figure ( figure 2 C) Marked by the black arrow (5000 times magnification), cell membrane rupture, mitochondrial swelling, and endoplasmic reticulum vesicle expansion can be seen.

Embodiment 3

[0252] Example 3 ɑ-NETA can cause ovarian cancer cells to secrete IL-18

[0253] In this experiment, ELISA kit was used to detect IL-18 in the supernatant of A2780 cells treated with ɑ-NETA.

[0254] First, use IL-18 standard substance (purchased from China Lianke) to carry out the establishment of standard curve (formula y=1396.6x^ 2 +1620.1x+128.73,R^ 2 = 0.9949). The A2780 cell supernatant treated with ɑ-NETA was used as the treatment group, and the cell supernatant treated with the same concentration of vehicle was used as the control group.

[0255] The results showed that there was IL-18 secretion in the treatment group (P image 3 A). By calculation, the concentration of ɑ-NETA is about 25 μg / ml (6.75*10 -2 μmol / ml) the concentration of IL-18 in the supernatant of A2780 cells after treatment was about 293.6pg / ml.

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Abstract

The present invention relates to application of alpha-NETA in the induction of cell death. In particular, the alpha-NETA has a very good death-inducing effect on tumor cells, particularly tumor cells of the reproductive system and the digestive system. The alpha-NETA can induce cells to rapidly die in a pyroptosis manner in a short time, and the required dosage is very low and is tens of times different from the dosage of conventional alpha-NETA.

Description

technical field [0001] The present invention relates to the field of cell death induction, specifically, the present invention relates to the application of α-NETA in inducing cell death, especially tumor cell death. Background technique [0002] Cell death refers to the irreversible cessation of life phenomena and the end of life. Cell death occurs in normal living tissues and is necessary to maintain tissue function and shape. The external factors often cause abnormal cell death. [0003] The result of cell death is mostly cell termination. Whether it is the programmed death of normal cells or abnormal cell death caused by external factors, cell death is closely related to the metabolism and health of the body. In fact, once the balance mechanism of cell death and regeneration in the normal body is broken, it will bring various pathological states to the body. For example, a tumor is a cell that loses its ability to undergo normal apoptosis and thus becomes a malignant ...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K31/567A61P35/00
CPCA61K31/567A61K45/06A61P35/00
Inventor 乔联桥席晓薇
Owner 乔联桥
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