Kit for quickly detecting expression quantities of related genes of sorafenib chemotherapeutic medicament

A kit and expression technology, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, fluorescence/phosphorescence, etc., can solve the problems of low patient survival rate and low patient diagnosis rate, and achieve simple and safe operation, fake Effect of low positives and high accuracy

Inactive Publication Date: 2012-08-01
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology

As far as the current situation of liver cancer in my country is concerned, not only the diagnosis rate of patients is very low, but also the survival rate of patients with advanced disease is also very low, almost less than 10%.
[0006] At present, there is no literature report on kits for detecting the expression of KDR and PDGFRA mRNA

Method used

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  • Kit for quickly detecting expression quantities of related genes of sorafenib chemotherapeutic medicament
  • Kit for quickly detecting expression quantities of related genes of sorafenib chemotherapeutic medicament
  • Kit for quickly detecting expression quantities of related genes of sorafenib chemotherapeutic medicament

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Embodiment 1

[0035] Embodiment 1: the preparation of kit of the present invention

[0036] (1) The kit of the present invention consists of the following:

[0037] ①Trizol: rapid extraction of cancer tissue RNA;

[0038] ② First strand cDNA synthesis kit (RT-PCR); including 25mmol / L MgCl 24μl, 10× reverse transcriptase buffer 2μl, 10mmol / L dNTP 2μl, RNase inhibitor 0.5μl, Oligo(dT) 15 0.5μg, AMV reverse transcriptase 15U, RNase-free deionized water;

[0039] ③ Primers: including target genes KDR, PDGFRA, internal reference gene GAPDH and fluorescent probes, as follows:

[0040] KDR gene primer sequence:

[0041] KDR gene upstream primer sequence is: 5'-CTGACGATTATGGAAGTGAGTGAAA-3';

[0042] KDR gene downstream primer sequence is: 5'-AAATGGGATTGGTAAGGATGACA-3';

[0043] KDR gene Taqman fluorescent probe: FAM 5'-AGACACAGGAAATTAC-3'TAMRA;

[0044] PDGFRA gene primer sequence:

[0045] The upstream primer sequence of PDGFRA gene is: 5'-CCTCATGGACGACCACCAA-3';

[0046] The downstream p...

Embodiment 2

[0056] Embodiment 2. detect the expression level of KDR, PDGFRA mRNA with the kit prepared in embodiment 1

[0057] Take the detection results of 30 cases of liver cancer paraffin section specimens as an example.

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Abstract

The invention relates to the technical field of biology. High expression of kinase insert domain receptors (KDR) and platelet-derived growth factor receptors alpha (PDGFRA) can be viewed in multiple tumor tissues; and sorafenib directly inhibits tumor growth by inhibiting the activities of the KDR and the PDGFR and inhibiting a recombinant activated factor/ methyl ethyl ketone/ extracellular signal-regulated kinase (RAF/ MEK/ ERK) signal transduction pathway, and blocks the formation of new tumor vessels by inhibiting vascular endothelial growth factors (VEGF) and platelet-derived growth factors (PDGF), so that the sorafenib achieves double inhibiting and multi-target blocking anti-hepatic cell carcinoma (HCC) effects. Detection of KDR and PDGFR mRNA levels can assist doctors in forecasting the curative effect of medicaments and the clinical outcome of patients, and has important clinical significance. The invention aims to provide a kit capable of quickly, conveniently, sensitively and specifically detecting expression quantities of related genes KDR and PDGFRA of a sorafenib chemotherapeutic medicament. According to the kit, the KDR and PDGFR mRNA levels are detected by adopting a fluorescent quantitative polymerase chain reaction (PCR) technology with high sensitivity and specificity, so that the sensitivity and the specificity are remarkably improved; and the kit is quick in detection and high in flux, and can finish the detection in 3 to 4 hours.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting the expression levels of Sorafenib chemotherapeutic drug-related genes KDR and PDGFRAmRNA by using fluorescent quantitative PCR technology. Background technique [0002] Hepatocellular carcinoma (HCC) is one of the most common clinical malignant tumors. According to the latest statistics, there are more than 600,000 newly diagnosed liver cancer patients in the world every year, of which nearly 50% occur in China, and the incidence of liver cancer is still constant increase. Currently, hepatocellular carcinoma (HCC) has become a fatal disease due to its strong aggressiveness and lack of systematic and effective treatment options. As far as the current situation of liver cancer in our country is concerned, not only the diagnosis rate of patients is very low, but also the survival rate of patients with advanced disease is also very low, almost less than 10%. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 刘辉汪珍光顾方明周伟平吴孟超
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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