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Method for applying pulse electric field combined carbon nanotube to facilitation of cancer cell apoptosis

A carbon nanotube, pulsed electric field technology, applied in biochemical equipment and methods, electric/wave energy treatment enzymes, and microbial assay/inspection, etc., can solve problems such as cell survival rebound, tissue damage, poor targeting, etc.

Pending Publication Date: 2018-11-23
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Needle-tip electrodes will create puncture points in the outer skin and cause tissue damage; while flat-plate electrode therapy has poor targeting, and high field strength is required to achieve the same therapeutic effect, and these strong electric fields will cause damage or even death to normal tissue cells
[0006] (2) After a period of time when pure electric pulses or carbon nanotubes act on cancer cells, the survival rate of cells rebounds significantly, and the effect of promoting apoptosis is not ideal

Method used

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  • Method for applying pulse electric field combined carbon nanotube to facilitation of cancer cell apoptosis
  • Method for applying pulse electric field combined carbon nanotube to facilitation of cancer cell apoptosis
  • Method for applying pulse electric field combined carbon nanotube to facilitation of cancer cell apoptosis

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Embodiment 1

[0040] The application of pulsed electric field combined with carbon nanotubes in the promotion of cancer cell apoptosis in this embodiment, in which the MTT method was used to detect the survival rate of cells in each treatment group, the specific implementation steps are as follows: Weigh an appropriate amount of carbon nanotubes, dissolve them in mPEG-pyrene (5mg / mL), sonicate at room temperature for 6h, centrifuge at 10000g for 1h, take the supernatant, and put the precipitate in an oven to dry and weigh to calculate the mass of dissolved carbon nanotubes. The supernatant was centrifuged at 4,000 rpm for 30 min with a 100 kDa ultrafiltration tube to remove excess mPEG-pyrene, and washed twice with double distilled water. The concentration is calculated according to the mass of the precipitated carbon nanotubes obtained by drying and the volume of the finally collected carbon nanotube solution. Digest the normally cultured human colon cancer cell line HCT116 cells with tryp...

Embodiment 2

[0042] The application of pulsed electric field combined with carbon nanotubes in promoting the apoptosis of cancer cells in this embodiment, wherein the AnnexinⅤ-FITC / PI double staining method is used to detect the apoptosis of arterial cells, the specific implementation steps are as follows: preparation of carbon nanotubes-cell suspension , the experimental grouping and pulse electric field related parameters are the same as in Example 1. After pulse treatment, take an appropriate amount of cells and culture them in normal medium in a 6-well plate. After 3 hours, collect the cells and resuspend the cells once with pre-cooled 1×PBS (4°C), centrifuge at 2000 rpm for 5-10 min, wash the cells, and add 300 μL of ×Binding Buffer to suspend the cells, then add 5 μL of Annexin V-FITC to each tube, mix well, incubate at room temperature for 15 minutes in the dark, then add 5 μL of PI for staining, and finally analyze the level of apoptosis by flow cytometry. The result is as Figure...

Embodiment 3

[0044] The application of pulsed electric field combined with carbon nanotubes in the promotion of cancer cell apoptosis in this embodiment, wherein the growth and proliferation ability of arterial cells is detected by cell cloning experiment, the specific implementation steps are as follows: carbon nanotube-cell suspension preparation, experimental grouping and pulse The parameters related to the electric field are the same as those in Example 1. After the pulse action, the cell suspension was diluted, and each group of cells was seeded in a 6-well plate at a density of 200 cells per well, and rotated gently to make the cells evenly dispersed. Set up 3 replicates for each treatment, placed at 37°C, 5% CO 2 and saturated humidity in a cell incubator for 2 to 3 weeks, during which the cell growth was closely observed. When colonies visible to the naked eye appeared in the Petri dish, the culture was terminated. Discard the supernatant, wash carefully with PBS twice, add 4% pa...

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Abstract

The invention discloses a method for applying a pulse electric field combined carbon nanotube to facilitation of cancer cell apoptosis, and relates to application of the pulse electric field combinedcarbon nanotube. The method comprises the following steps: preparing a cell suspension; preparing deionized dispersion liquids of different concentrations of three kinds of carbon nanotubes; uniformlymixing the carbon nanotube deionized dispersion liquids with the cell suspension to obtain a carbon nanotube-cell suspension; adding the cell suspension into an electrode cup to set a blank control group and a pure electric pulse treatment group; adding the carbon nanotube-cell suspension into the electrode cup for setting a pure carbon nanotube treatment group and a pulse electric field combinedcarbon nanotube treatment group; applying a pulse electric field to a positive electrode plate and a negative electrode plate of the electrode cup in an experiment group being subjected to the pulsetreatment; measuring the cell survival rate of each treatment group by using an MTT method after the pulse treatment; analyzing the cell apoptosis of each treatment group through a flow cytometry by an Annexin V-FITC / PI double staining method after the pulse treatment; detecting the change of cell growth and multiplication capabilities through clone formation assay after the pulse treatment.

Description

technical field [0001] The invention relates to the application of pulsed electric field combined with carbon nanotubes, in particular to the application method of pulsed electric field combined with carbon nanotubes in promoting apoptosis of cancer cells. Background technique [0002] The application of pulsed electric fields in promoting cell death, that is, the use of electric fields with pulse widths of milliseconds and microseconds and field strengths from tens of V / cm to tens of kV / cm to act on cell tissues, using the unique bioelectric effect of pulsed electric fields , to achieve the purpose of cell death. The mechanism of traditional millisecond pulse or microsecond pulse electric field to promote the death of cancer cells is to change the function of cancer cells through reversible or irreversible electroporation, and at the same time induce the body's apoptosis effect, anti-vascular and lymphatic metastasis effects, and destroy the living conditions of cancer cell...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12Q1/02
CPCC12N13/00G01N33/5011
Inventor 章幼玉汪斌毛铮陆楠钟渊福柳清伙
Owner XIAMEN UNIV
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