Applications of phosphodiesterase PDE3A in determination of tumor treatment effect of Anagrelide

A technology for tumors and tumor cells, applied in disease diagnosis, microbial measurement/testing, biochemical equipment and methods, etc., can solve problems such as lack of diagnostic methods

Inactive Publication Date: 2017-11-03
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that anagrelide or its pharmaceutically acceptable preparations have new uses in inhibiting tumors, but anagrelide does not have a good effect on all types of tumors, and there is still a lack of clear diagnostic methods to guide anagrelide. Gray's Medication

Method used

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  • Applications of phosphodiesterase PDE3A in determination of tumor treatment effect of Anagrelide
  • Applications of phosphodiesterase PDE3A in determination of tumor treatment effect of Anagrelide
  • Applications of phosphodiesterase PDE3A in determination of tumor treatment effect of Anagrelide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0181] Detection of PDE3A Protein Expression in Tumor Cell Lines

[0182] In order to study the expression of PDE3A protein in tumor cells, the expression level of PDE3A protein was detected on the cell lines listed in the general materials by Western blotting.

[0183] The result is as figure 1 As shown, PDE3A protein is positively expressed in cell lines such as SMMC7721, FHCC98, H4, HeLa, Bel7404, A498, SW1116, and negatively expressed in other 20 cell lines.

[0184] Further, RT-PCR was used to detect the PDE3A mRNA level of some cell lines; RQ: ​​the quantitative result of relative β-actin.

[0185] The result is as image 3 As shown, there was PDE3A mRNA expression in SMMC7721, FHCC98, H4, HeLa, Bel7404, A498 and SW1116, but there was almost no PDE3A mRNA expression in H522 and SKOV3, which was consistent with the detection results of Western blot.

Embodiment 2

[0187] Drug anti-tumor activity test

[0188] Utilize the Anagrelide (Anagrelide) that concentration is 1 μ M to carry out activity experiment to the cell line in embodiment 1, according to cell viability, cell line is divided into three classes: sensitive (S), moderately sensitive (M), insensitive ( R), and further determine the IC50 value of anagrelide inhibiting each cell line.

[0189] The result is as figure 2 As shown, there were 7 sensitive cells and 20 insensitive cells in the test cell lines ( figure 2 ). Table 1 shows the IC50 values ​​of anagrelide-sensitive cell lines, among which, the cell lines SMMC7721, FHCC98, H4, and HeLa positively expressing PDE3A are sensitive cells of anagrelide, and anagrelide has an excellent inhibitory effect on it. The IC50 values ​​were all less than 30nM; the cell lines Bel7404, A498 and SW1116 positively expressing PDE3A were moderately sensitive to anagrelide, and anagrelide had a lasting inhibitory effect on it, with IC50 val...

Embodiment 3

[0194] Effect of PDE3A protein knockout on the growth of sensitive cells

[0195] Examples 1 and 2 illustrate the relationship between PDE3A protein and growth inhibition from the perspective of compounds inhibiting protein activity, but the compounds will cause differences in cell selectivity due to their different structures and different modes of action with the target. Therefore, after obtaining the PDE3A protein cell expression profile data, the siRNA interference technology was used to investigate the effect of knocking out the PDE3A protein on cell growth. Experiments were performed with two pde3a siRNAs (with different efficiencies), and cells were plated on RTCA plates 48 hours after the siRNAs were transfected for long-term growth monitoring.

[0196] The result is as Figure 4 Shown: the expression level of PDE3A protein ( Figure 4 A) is related to the speed of cell growth ( Figure 4 B) There is a positive correlation, indicating that PDE3A protein plays an imp...

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Abstract

The present invention relates to applications of phosphodiesterase PDE3A and a gene thereof in determination of the tumor treatment effect of Anagrelide, and specifically provides use of a PDE3A gene sequence, a protein, or a specific antibody against PDE3A protein in preparation of a diagnostic reagent or a diagnostic kit, wherein the diagnostic reagent or kit is used for (a) determining the tumor treatment effect of anagrelide, and / or (b) determine whether tumor patients can be treated with anagrelide. The invention further provides a corresponding test kit.

Description

technical field [0001] The invention relates to the field of tumor molecular detection. More specifically, it relates to the application of PDE3A in judging the effect of anagrelide in treating tumors. Background technique [0002] The occurrence and development of tumors are the result of multiple genes and multiple environmental factors. The diversity of genetic backgrounds of tumor patients determines the complexity of tumor treatment, and also indicates that individualized treatment is required for this disease. "Targeted therapy" that uses targeted technology to precisely deliver drugs to tumor lesions and "targeted therapy" that utilizes tumor-specific gene mutations and gene functions are currently hot spots in cancer drug research. Molecular target therapy is aimed at specific key molecules in different tumor cells that promote tumor growth and survival, and achieve anti-tumor effects that inhibit tumor cell growth or promote apoptosis. Representative target drugs ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574
CPCC12Q1/6886G01N33/57484G01N33/6854G01N2333/916C12Q2600/106C12Q2600/158G01N33/574G01N2800/52
Inventor 俞强刘珏妤
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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