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Light-up fluorescent probe for identifying and detecting liver cancer cells

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

Active Publication Date: 2018-08-24
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Abstract
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  • Claims
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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

Method used

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  • Light-up fluorescent probe for identifying and detecting liver cancer cells
  • Light-up fluorescent probe for identifying and detecting liver cancer cells

Examples

Experimental program
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Effect test

Embodiment 1

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

[0032] (2) Dilute solution a with high-sugar DMEM medium and prepare them to a concentration of 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 fluorescent probes with different concentrations are obtained;

[0033] (3) Take six 96-well cell culture plates, take one row of wells in each 96-well cell culture plate as the blank control group, and the other wells as the experimental group; pass HepG2cell to three 96-well cell culture plates, and pass L02cell In the other three 96-well cell culture plates, each well of the experimental group contained 100 μL of cell suspension (about 5000 cells), and each well of the control group contained 100 μL of cell-free high-sugar DMEM medium; six 96-well cell culture plate placed at 37℃, CO 2 After culturing in a 5% cell culture incubator for 24 hours, add 100μL of high-sugar DM...

Embodiment 2

[0036] (1) Dissolve ZZ-HPB-NC in DMSO to make 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 used to identify and detect liver cancer cells;

[0038] (3) Three kinds of human liver cancer cells (HepG2cell, Hep3Bcell, SMMC7721cell) and two kinds of normal human liver cells (L02cell, QSG7701cell) that have been covered with 80% to 90% of the bottom of the petri dish were used with trypsin with a mass fraction of 0.05% (Diluted with 0.25% mass fraction of commercial trypsin solution with PBS) Digest and pass into cell culture dishes (the number of cells in each cell culture dish is about 1×10 5 Pcs) and placed in 37℃, CO 2 Cultivate in a 5% cell culture incubator for 24 hours; then aspirate the medium, and add 1 mL of solution b to each cell culture dish. After standing for 5 minutes for staining, first aspirate the medium, and then rinse with PBS 3 times to remove For the ...

Embodiment 3

[0041] (1) Dissolve ZZ-HPB-NC in DMSO to make 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 used to identify and detect liver cancer cells;

[0043] (3) Pass HepG2cell and L02cell into cell culture dishes at a quantitative ratio of 10:1, 1:1, and 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℃, CO 2 Cultivate for 24 hours in a cell culture incubator with a volume content of 5%, in which there are 4 parallel cell culture dishes for each number ratio; then aspirate the medium, and add 0.3 mL of solution b to each cell culture dish, and let stand for 10 minutes for staining Then, first aspirate the medium, and then rinse with PBS 3 times to remove the free solution b, and then add 2 mL of high-sugar DMEM medium and place the cell culture dish under a fluorescent microscope to observe the intrac...

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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 lighting type fluorescent probe containing a hexaphenylbutadiene derivative containing methyl 2-cyanoacetate for rapid identification and detection of liver cancer cells in situ, which belongs to biological fluorescent labeling and cell biology field. Background technique [0002] Liver cancer is a common malignant tumor disease of the liver, divided into two categories: primary and secondary. Primary liver cancer has a very high incidence in my country, and it is a malignant tumor that is quite harmful to human health; secondary liver cancer is also called metastatic liver cancer, which is generally common in the biliary tract, colorectal, stomach, pancreas, uterus, ovary, and breast Liver metastasis of malignant tumors in other organs. Whether it is primary liver cancer or secondary liver cancer, it has become the number one killer of human health in the world. Because the early symptoms of liver cancer are not ob...

Claims

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Application Information

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Patent Type & Authority Applications(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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