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A light-up fluorescent probe for identification and detection of liver cancer cells

A technology for fluorescent probes and liver cancer cells, applied in fluorescence/phosphorescence, measuring devices, and material analysis through optical means, can solve the problems of shortening the early diagnosis time of liver cancer, high cost of inspection and diagnosis, and few early diagnosis methods. Achieve the effects of shortening the time of early diagnosis, reducing physical and mental suffering, and reducing economic burden

Active Publication Date: 2020-09-11
BEIJING INSTITUTE OF TECHNOLOGYGY
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Problems solved by technology

[0004] Aiming at the problems that there are few early diagnosis methods for liver cancer at present, they are easily affected by operator’s human factors and need to use preparations with toxic side effects on normal tissues and organs of the human body, and the cost of inspection and diagnosis is high. The purpose of the present invention is to provide a method for identification and diagnosis A light-emitting fluorescent probe for detecting liver cancer cells. The main component of the fluorescent probe is 2,2'-((((1Z,3Z)-1,2,3,4-tetraphenyl-1,3-butyl Diene-1,4-)bis(4,1-phenyl))divinyl-(2-cyano)acetic acid methyl ester (English abbreviation: ZZ-HPB-NC), this component is only specific to liver cancer cells It has a fluorescent labeling effect, but has no fluorescent labeling effect on normal liver cells, so that it can effectively distinguish liver cancer cells from normal liver cells; at the same time, it is easy to use the fluorescent probe as a fluorescent indicator to identify and detect liver cancer cells. Low cost, less toxic and side effects, and can shorten the early diagnosis time of liver cancer, can be used as a potential early liver cancer diagnostic reagent and a fluorescent indicator for clinical liver cancer surgery

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  • A light-up fluorescent probe for identification and detection of liver cancer cells
  • A light-up fluorescent probe for identification and detection of liver cancer cells

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Experimental program
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Effect test

Embodiment 1

[0031] (1) ZZ-HPB-NC was dissolved in DMSO to prepare 10 -3 mol / L solution a;

[0032] (2) Dilute solution a with high-sugar DMEM medium, and prepare the concentration as 1×10 -4 mol / L, 5×10 -5 mol / L, 1×10 -5 mol / L, 5×10 -6 mol / L, 1×10 -6 mol / L, 1×10-7 mol / L solution b 1 , b 2 , b 3 , b 4 , b 5 , b 6 , that is, six different concentrations of fluorescent probes were obtained;

[0033] (3) Take six 96-well cell culture plates, take a row of wells in each 96-well cell culture plate as the blank control group, and the remaining wells as the experimental group; pass HepG2cell to three 96-well cell culture plates, and L02cell passage In another three 96-well cell culture plates, 100 μL of cell suspension (about 5000 cells) in each well of the experimental group, and 100 μL of cell-free high-sugar DMEM medium in each well of the control group; six The 96-well cell culture plate was placed at 37°C, CO 2 After cultivating in a cell culture incubator with a volume content ...

Embodiment 2

[0036] (1) ZZ-HPB-NC was dissolved in DMSO to prepare 10 -3 mol / L solution a;

[0037] (2) Add high-sugar DMEM medium to solution a to obtain a concentration of 10 -6 mol / L solution b, which is a fluorescent probe for identifying and detecting liver cancer cells;

[0038] (3) Three kinds of human hepatoma cells (HepG2cell, Hep3Bcell, SMMC7721cell) and two kinds of human normal hepatocytes (L02cell, QSG7701cell) that had covered 80% to 90% of the bottom of the culture dish were respectively treated with 0.05% trypsin (diluted commercial trypsin solution with a mass fraction of 0.25% in PBS) digested and passaged into cell culture dishes (the number of cells in each cell culture dish is about 1×10 5 ), and placed at 37°C, CO 2 Culture in a cell culture incubator with a volume content of 5% for 24 hours; then aspirate the medium, and add 1mL of solution b to each cell culture dish, let it stand for 5 minutes for staining, first aspirate the medium, and then rinse with PBS 3 ti...

Embodiment 3

[0041] (1) ZZ-HPB-NC was dissolved in DMSO to prepare 10 -3 mol / L solution a;

[0042] (2) Add high-sugar DMEM medium to solution a to obtain a concentration of 10 -6 mol / L solution b, which is a fluorescent probe for identifying and detecting liver cancer cells;

[0043] (3) Passage HepG2cell and L02cell to the cell culture dish according to the ratio of 10:1, 1:1, 1:10 respectively (the total number of two kinds of cells in each dish is about 1×10 5 ), and place the cell culture dish at 37°C, CO 2 Cultivate in a cell culture incubator with a volume content of 5% for 24 hours, and there are 4 parallel cell culture dishes for each quantity ratio; then suck out the medium, add 0.3mL solution b to each cell culture dish, and let stand for 10 minutes for staining Finally, suck out the culture medium first, then rinse with PBS 3 times to remove the free solution b, then add 2 mL of high-sugar DMEM medium, place the cell culture dish under a fluorescence microscope to observe th...

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Abstract

The invention relates to a light-up fluorescent probe for identifying and detecting liver cancer cells, and belongs to the field of bioluminescent labeling and cell biology. The fluorescent probe is amixed solution prepared from ZZ-HPB-NC, dimethyl sulfoxide and a high glucose DMEM medium; whether being used in a single cell culture system or a co-culture system of the liver cancer cells and normal liver cells, the fluorescent probe only has a specific light-up fluorescent labeling effect on the liver cancer cells, and has no fluorescent labeling effect on the normal liver cells, so that thelight-up fluorescent probe can effectively screen the liver cancer cells and the normal liver cells; meanwhile, the light-up fluorescent probe as a fluorescent indicator for identifying and detectingthe liver cancer cells is simple, low in cost, and small in toxic and side effects, can shorten the early diagnosis time of liver cancer, and can be used as a potential early liver cancer diagnostic reagent and a clinical liver cancer surgical fluorescent indicator.

Description

technical field [0001] The invention specifically relates to a light-up fluorescent probe containing a hexaphenylbutadiene derivative containing methyl 2-cyanoacetate for in-situ rapid identification and detection of liver cancer cells, which belongs to the field of bioluminescence labeling and cell biology field. Background technique [0002] Liver cancer is a common malignant tumor of the liver, which can be divided into two categories: primary and secondary. Primary liver cancer has a very high incidence in my country and is a malignant tumor that poses a considerable hazard to human health; secondary liver cancer, also known as metastatic liver cancer, is commonly found in the biliary tract, colorectum, stomach, pancreas, uterus, ovary, and breast. Liver metastases from other organ malignancies. Whether it is primary liver cancer or secondary liver cancer, it has become the number one killer of human health in the world. Since the early symptoms of liver cancer are not...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428
Inventor 陈笛笛毛慧灵张亚会董宇平
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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