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Nano-fluorescent probe targeting pancreatic cancer circulating tumor cells

A nano-fluorescent probe and tumor cell technology, applied in the field of "ratio-type" nano-fluorescent probes of intracellular hydrogen peroxide, can solve the problems of reduced expression and disappointing results

Active Publication Date: 2019-01-22
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, due to the heterogeneity of tumor cells in pancreatic cancer, the expression of epithelial markers such as EpCAM and CK19 is very likely to decrease during the epithelial-mesenchymal transition (EMT) of the cells.
Therefore, there are limitations in the application of EpCAM and CK19 antibodies to capture enrichment and identification of CTCs, respectively, and the results were disappointing in the application of CTCs in capturing pancreatic cancer

Method used

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  • Nano-fluorescent probe targeting pancreatic cancer circulating tumor cells
  • Nano-fluorescent probe targeting pancreatic cancer circulating tumor cells
  • Nano-fluorescent probe targeting pancreatic cancer circulating tumor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Synthesis of CD44-targeted nano-fluorescent probes and tumor-targeted detection

[0035] figure 1 For the synthesis of hyaluronic acid-naphthalene imine-borate graft polymer, 160mg (0.4mmol) sodium hyaluronate was dissolved in 50mL ultrapure water, and vigorously stirred for 2h to fully dissolve. Subsequently, 117.8 mg (0.4 mmol) DTMMM was added to activate the carboxyl groups on the hyaluronic acid for 30 min. Finally, 43.9 mg (0.12 mmol) NP-BE dissolved in DMSO was added dropwise to the above mixture, and reacted at room temperature for 24 h in the dark. After the reaction, the above reaction mixture was dissolved in DMSO / H with a molecular weight cut-off of 14kDa in a dialysis bag. 2 O (1:4v / v) mixed solvents were dialyzed for two days, and dialyzed in pure water for two days. The aqueous solution purified by dialysis was freeze-dried to obtain the HA-NP-BE grafted polymer. HA-NP-BE grafted polymers are amphiphilic and can self-assemble in aqueous solu...

Embodiment 2

[0045] Example 2. Identification of circulating tumor cells in patients with pancreatic cancer

[0046] HA-NP-BE nanofluorescent probe can target endogenous H in pancreatic cancer cells 2 o 2 , and has fast responsiveness, can effectively distinguish pancreatic cancer tumor cells from normal cells, and apply it to the detection of circulating tumor cells in pancreatic cancer patients in auxiliary clinical practice. The Ethics Committee of Shanghai Jiaotong University approved the experiment. The peripheral blood of clinical pancreatic cancer patients was collected, and circulating tumor cells were enriched by negative selection. Firstly, the erythrocytes were removed by red blood cell lysate, most of the leukocytes in the blood were removed by CD45 antibody-labeled magnetic beads, and then the enriched CTCs were detected with HA-NP-BE nano-fluorescence probe and CK19-Alexa Fluor 647 respectively. stained, and finally observed and counted under a confocal laser microscope. ...

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Abstract

The invention provides a nano-fluorescent probe targeting pancreatic cancer circulating tumor cells. The probe comprises H2O2 responsive fluorescent small molecules and polymers targeting pancreatic cancer surface markers, and the H2O2 responsive fluorescent small molecules and the polymers targeting the pancreatic cancer surface markers have opposite hydrophilicity and hydrophobicity. Through thehigh expression of CD44 antigen specificity binding of hyaluronic acid and the pancreatic cancer tumor surface, the HA-NP-BE nano-fluorescent probe can tragetingly enter pancreatic cancer cells to detect H2O2 in the cells and image H2O2, rapid responsiveness is achieved, the pancreatic cancer tumor cells and normal cells can be effectively distinguished, CTC in pancreatic cancer tumor patients isdetected, and early disgnosis of pancreatic cancer is facilitated.

Description

technical field [0001] The invention belongs to the field of biology, and more specifically relates to a "ratio-type" nanometer fluorescent probe targeting intracellular hydrogen peroxide in circulating tumor cells of pancreatic cancer. Background technique [0002] At present, the detection of circulating tumor cells (CTCs) mainly includes two parts: enrichment and identification. Due to the characteristics of small amount, heterogeneity and deformability of CTCs, many methods are currently available for the detection of CTCs. Enrichment methods include physical methods based on cell density, size, dielectric properties, etc., immunological methods based on cell surface specific antigens including epithelial adhesion molecule (EpCAM) and EGFR, and negative screening methods for removing red blood cells and white blood cells; Identification methods include flow cytometry, immunofluorescence, fluorescence in situ hybridization, reverse transcription PCR and next-generation s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06C08B37/08
CPCC08B37/0072C09K11/025C09K11/06C09K2211/1088C09K2211/1491G01N21/6428
Inventor 李培勇李纯婷朱新远
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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