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A novel screening method for anticancer drugs

A screening method and anti-cancer drug technology, applied in the field of molecular biology, can solve the problems of not being able to understand the mechanism of anti-cancer action, being easily affected by human error, consuming time and funds, etc., and achieving easy promotion, simple method, and few reagents Effect

Inactive Publication Date: 2015-09-30
THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method requires manual counting of cells, which is susceptible to human error
[0005] 3. Nuclide precursor incorporation experiment ( 3 H-Tdr Assay), this method requires less cells, so needle aspiration samples can be used, but this method only measures 5% to 10% of the cells in the sample, thymine and normal cells will affect the measurement results, and there is radioactivity Pollution
[0013] Disadvantages of the prior art: At present, all drug sensitivity tests can only screen out drugs that effectively inhibit tumor growth, but cannot understand the anti-cancer mechanism of the drug that is screened out. If the mechanism of drug action is not understood, it will greatly affect the application of it to Clinical research; to understand the mechanism of drug action, other experiments are still needed for research, which consumes a certain amount of time and money

Method used

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  • A novel screening method for anticancer drugs
  • A novel screening method for anticancer drugs
  • A novel screening method for anticancer drugs

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Embodiment

[0035] A novel screening method for anticancer drugs, the key of which is to include the following steps:

[0036] (1) Preparation of TTLL12 overexpression cell line

[0037] 1) PCR amplification of TTLL12 gene fragment

[0038] Self-designed primers, sense strand 5'-CGGGATCCCCGATGGACTACCACGAGGA-3', antisense strand 5'-CGGGATCCTAGCTACAGGACGCCCCCG-3', amplified the TTLL12 gene fragment using TTLL12 cDNA as a template. The size of the target gene fragment is 2818bp, the composition of PCR reaction solution: Ex Taq enzyme (5u / μL) 0.15μL, 10×PCR buffer 2.5μL, MgCL 2 (25mmol / L) 2μL, dNTP (each 2.5mmoL / L) 2μL, upstream primer (10mol / L) 1.0μL downstream primer (10mol / L) 1.0μL, template DNA 4.5μL, add sterilized pure water to 25μL. PCR cycle parameters: 94°C for 5min, 94°C for 50s, 55°C for 30s, 72°C for 30s, 35cycle, 72°C for 8min. Identification of PCR products: Separation and identification on 1% agarose gel electrophoresis. see results figure 2 .

[0039] 2) Insert the TTLL...

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Abstract

The invention discloses a novel screening method of anticancer medicaments. The novel screening method is characterized by comprising the followinh steps of: (1) preparing TTLL12 overexpressed Hep-2 cell lines; (2) selecting positive control and negative control medicaments; (3) obtaining the optimum concentration of nitrotyrosine and the optimum diluted concentrations of medicaments to be detected; and (4) screening the medicaments, finally checking readings and comparing. The novel screening method of the anticancer medicaments disclosed by the invention has the characteristics that the method is simple, a user only needs to master a cell culture and Elisa (Enzyme-linked Immuno Sorbent Assay) technology and then masters the law, the method is fast and cheap, the method is few in reagents and variation factors, stable, high in sensitivity, good in specificity, easy to popularize and capable of screening in a large scale. Furthermore, through the experiment, whether the medicaments to be detected have anti-cancer effects and anticancer effect mechanisms of the medicaments to be detected simultaneously can be obtained, so that the speed of clinically applying the medicaments to be detected is greatly accelerated, and more lives of patients are expected to be saved.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a novel screening method for anticancer drugs. Background technique [0002] Malignant tumors are a large class of diseases that seriously threaten human life and health. Chemotherapy, as a systemic treatment, has introduced folic acid antagonists as antitumor drugs into the treatment of leukemia since 1948, and has an irreplaceable position in the comprehensive treatment of malignant tumors. However, due to the limited types of current drugs and the emergence of Therefore, the curative effect of chemotherapy is still not satisfactory, and we still need to continue to find more effective drugs. There are many screening methods for anticancer drugs, such as: [0003] 1. Human Tumor Cell Cloning Assay (HTCA), the advantage of this method is: it is sensitive and can directly evaluate cell proliferation and death. But its disadvantages are: low detection rate ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/15
Inventor 李雅冬
Owner THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIVERSITY
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