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Method for detecting effect of Coptis chinensis alkaloid monomer on tumor cells

A technology of coptis alkaloids and tumor cells, which is applied in the field of detecting the effect of coptis alkaloid monomers on tumor cells, can solve the problems of inability to obtain the intuitive dynamic process of drug action, poor sensitivity time point observation, cumbersome operation, etc., to avoid cell rupture , Small size, saving manpower and material resources

Inactive Publication Date: 2014-11-26
THE FIFTH MEDICAL CENT OF CHINESE PLA GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods use the changes in cell proliferation and viability before and after administration as evaluation indicators, require specific labeling and processing, are cumbersome to operate, have poor sensitivity and can only be observed at specific time points, and cannot obtain an intuitive dynamic process of drug action

Method used

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  • Method for detecting effect of Coptis chinensis alkaloid monomer on tumor cells
  • Method for detecting effect of Coptis chinensis alkaloid monomer on tumor cells
  • Method for detecting effect of Coptis chinensis alkaloid monomer on tumor cells

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1: The present invention uses a real-time cytometer to detect the effect of Coptis Rhizoma alkaloid monomers on tumor cells. The tumor cells are specifically liver cancer cells HepG2; Alkaloids, coptisine, jatrorrhizine, epiberberine, the method of the present invention can simultaneously monitor the effects of different concentrations and various alkaloid monomers on liver cancer cells HepG2, and the first example is for different concentrations of berberine The effect of alkali liquid on liver cancer cells HepG2, the real-time cell analyzers used in this embodiment are all DP type real-time cell analyzers.

[0047] 1. Instruments used in the test: RTCA DP (German Roche Company); E-Plate 16 detection plate (6x6 plates, German Roche Company); constant temperature incubator (5410 type, American Napco Company); refrigerated centrifuge (HC- 3018R, Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); electronic balance (AL-204, Mettler-Toledo Instruments (Sha...

Embodiment 2

[0061] Example 2: This example lists the effect of coptisine liquid on liver cancer cells HepG2 at different concentrations. The test equipment used is the same as that in Example 1. The reference substance of coptisine (purity greater than 98%) was purchased from Chengdu Mansi Special Biotechnology Co., Ltd.

[0062] The method for detecting the effect of coptisine liquid at different concentrations on liver cancer cell HepG2 is as follows: (1) prepare different concentrations of liver cancer cell HepG2 suspension and coptisine liquid; Monitor the cell growth index in the cell HepG2 suspension to determine the optimal cell concentration of the used liver cancer cell HepG2 suspension and the best administration time point for adding the drug solution when detecting the effect of coptisine liquid on the liver cancer cell HepG2; (3) Use the real-time cell analyzer to detect the growth index value of the liver cancer cell HepG2 before and after administration, and judge the effec...

Embodiment 3

[0069] Example 3: This example lists the effects of palmatine liquid on liver cancer cells HepG2 at different concentrations. The test equipment used is the same as that in Example 1. Palmatine reference substance (purity greater than 98%) was purchased from Chengdu Mann Ster Biotechnology Co., Ltd.

[0070] The method for detecting the effect of palmatine liquid at different concentrations on liver cancer cell HepG2 is as follows: (1) prepare different concentrations of liver cancer cell HepG2 suspension and palmatine liquid; The cell growth index in the palmatine suspension is monitored to determine the optimal cell concentration of the liver cancer cell HepG2 suspension used and the optimal administration time of adding the drug solution when detecting the effect of the palmatine liquid on the liver cancer cell HepG2 (3) Use the real-time cell analyzer to detect the tumor cell growth index value before and after administration, and judge the effect of the drug solution on t...

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Abstract

The invention discloses a method for detecting effect of a Coptis chinensis alkaloid monomer on tumor cells. The method comprises the following steps: (1) respectively preparing tumor cell suspensions and Coptis chinensis alkaloid monomer soups with different concentrations; (2) monitoring cell growth indexes in the tumor cell suspensions with different concentrations by means of a real time cell analyzer and determining the optimal cell concentration of the tumor cell suspensions used and the optimal feeding time point for adding the soup when the effect of the Coptis chinensis alkaloid monomer on tumor cells is detected; and (3) determining the tumor cell growth values before and after delivery by means of the real time cell analyzer and judging the effect of the soup on the tumor cells according to the cell growth index values. The method provided by the invention has the advantages that the method can reflect physiological statuses of growth, expansion, morphologic change and death of the tumor cells before and after delivery in real time, so that the method has high sensitivity and repeatability, and is free from marks and stable and accurate in detection result.

Description

technical field [0001] The invention belongs to the technical field of methods for detecting cell activity in organisms, in particular to a method for detecting the effect of coptis alkaloid monomers on tumor cells. Background technique [0002] Coptis Chinensis Franch (Coptis Chinensis Franch) is a commonly used traditional Chinese medicine. The main alkaloids contained in Coptis Chinensis are berberine, coptisine, jatrorrhizine, palmatine, etc. Studies in recent years have shown that berberine and coptisine have certain anti-tumor effects, and can inhibit and kill a variety of cancer cells, such as liver cancer HepG2 cells, lung cancer GLC-82 cells, gastric cancer MGC-803 cells, etc. The search for TCM monomers with anti-tumor activity has attracted more and more attention. To evaluate whether various TCM monomers have the effect of killing tumor cells depends on the detection of their anti-tumor activity. At present, traditional breakpoint detection methods are mainly us...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02
Inventor 鄢丹张乐乐马丽娜章从恩熊呤肖小河
Owner THE FIFTH MEDICAL CENT OF CHINESE PLA GENERAL HOSPITAL