A method for studying anticancer drugs by monitoring endocytosis in real time

Abstract

The invention provides a method for real-time monitoring of cell endocytosis to study anticancer drugs. The method for real-time monitoring of the endocytosis process provided by the present invention comprises the following steps: 1) adding anticancer drugs to make it act on the cells to be tested; 2) connecting epidermal growth factor on the cell membrane of the cells to be tested by a two-step labeling method -Quantum dot probe; 3) Using a fluorescence microscope to observe and photograph under the condition that the light intensity is less than 100 μW, and track the endocytosis process according to the quantum dot label; 4) Quantitatively analyze the photographed data to obtain a relative endocytosis curve. The method provided by the present invention achieves the study of drug effects by quantitatively analyzing the effect of anticancer drugs on endocytosis through real-time observation throughout the whole process, and also considers the influence of cell shape to standardize the endocytic transport data of different cells, completely It can realize horizontal comparison and provide a new quantitative research method for the research, development, screening and therapeutic effect of anticancer drugs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular, the invention relates to a method for researching anticancer drugs by monitoring cell endocytosis in real time. Background technique [0002] With the rapid development of life science research, bioluminescent technology has been widely used in cellular immunology, molecular biology, oncology and other fields. Fluorescence technology has become one of the important means of life science research. However, traditional fluorescent probes such as green fluorescent protein (GFP), Alexa Fluro, Cy3, Cy5, etc. have some insurmountable shortcomings, such as different degrees of fluorescence quenching, unstable light intensity, and cannot be applied to long-term observation experiments . However, many life processes are long-term and continuous changes, so a fluorescent probe with good stability is needed. [0003] Quantum dots (quantum dots, QDs), also known as semiconductor nanocrystals, a...

AI Technical Summary

Problems solved by technology

[0005] The current research strategy on cell endocytosis is to take pictures at different time points to obtain data, and find the effects of anticancer drugs through static pictures at several time points. Most of them are qualitative conclusions, and they are not specific to a specific cell.

When such results are compared horizontally, their credibility is reduced. More importantly, comprehensive, dynamic and real-time visible data of endocytic transport cannot be obtained.

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