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Gene combination used for guiding individual treatment of platinum medicines

A platinum-based drug and gene combination technology, applied in the field of genetics, can solve problems such as differences in platinum-based drugs

Inactive Publication Date: 2012-03-07
SHANGHAI BIOTECAN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The technical problem to be solved by the present invention is to provide a convenient and fast gene combination and its application to assist clinicians in formulating individualized drug regimens in view of the obvious differences in the curative effect of platinum-based drugs among different patients in the current tumor treatment process. method

Method used

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  • Gene combination used for guiding individual treatment of platinum medicines
  • Gene combination used for guiding individual treatment of platinum medicines
  • Gene combination used for guiding individual treatment of platinum medicines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 A method for identifying mutations in GSTP1

[0032] 1. Extract the genomic DNA of the patient's tumor tissue;

[0033] 2. Perform PCR amplification. The 20 μl PCR reaction system is as follows: 50-100 ng of genomic DNA of the tumor tissue to be tested, 0.125 μl of Taq enzyme, 1 μl of upstream and downstream primers (10 μM), 2 μl of dNTP (2.5 mM), 10 × PCR buffer (containing Mg 2+) 2 μl, and the rest was sterile distilled water; the PCR reaction conditions were: 95°C pre-denaturation for 2 min, followed by 95°C denaturation for 30 sec, 55°C annealing for 30 sec, 72°C extension for 30 sec, 40 cycles, and finally 72°C extension for 7 min, 4°C storage ;

[0034] 3. Gel electrophoresis analysis of PCR amplification products:

[0035] a. Rinse the electrophoresis tank and comb with distilled water. Put it on a level table and set up the comb. 1.5% agarose can be prepared for electrophoresis.

[0036] b. Put 50ml of 1×TAE electrophoresis buffer and 100ml into a...

Embodiment 2

[0055] Example 2 A method for identifying MRP2 mutations

[0056] 1. Extract the genomic DNA of the patient's tumor tissue;

[0057] 2. Perform PCR amplification. The 20 μl PCR reaction system is as follows: 50-100 ng of genomic DNA of the tumor tissue to be tested, 0.125 μl of Taq enzyme, 1 μl of upstream and downstream primers (10 μM), 2 μl of dNTP (2.5 mM), 10 × PCR buffer (containing Mg 2+ ) 2 μl, and the rest was sterile distilled water; the PCR reaction conditions were: 95°C pre-denaturation for 2 min, followed by 95°C denaturation for 30 sec, 64°C annealing for 30 sec, 72°C extension for 30 sec, 40 cycles, and finally 72°C extension for 7 min, 4°C storage ;

[0058] 3. Gel electrophoresis analysis of the PCR amplification product: the specific steps are the same as in Example 1.

[0059] The result is as figure 1 As shown, band 3 represents MRP2, and its PCR product is 414bp.

[0060] 4. Sequencing of PCR amplification products: the specific steps are the same as i...

Embodiment 3

[0062] Example 3 A method for identifying mutations in XRCC1-Exon6

[0063] 1. Extract the genomic DNA of the patient's tumor tissue;

[0064] 2. Perform PCR amplification. The 20 μl PCR reaction system is as follows: 50-100 ng of genomic DNA of the tumor tissue to be tested, 0.125 μl of Taq enzyme, 1 μl of upstream and downstream primers (10 μM), 2 μl of dNTP (2.5 mM), 10 × PCR buffer (containing Mg 2+ ) 2 μl, and the rest was sterile distilled water; the PCR reaction conditions were: 95°C pre-denaturation for 2 min, followed by 95°C denaturation for 30 sec, 58°C annealing for 30 sec, 72°C extension for 30 sec, and 40 cycles, and finally 72°C extension for 7 min, and 4°C storage ;

[0065] 3. Gel electrophoresis analysis of the PCR amplification product: the specific steps are the same as in Example 1.

[0066] The result is as figure 1 As shown, band 4 represents XRCC1-Exon6, and its PCR product is 330bp.

[0067] 4. Sequencing of PCR amplification products: the specifi...

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Abstract

The invention belongs to the gene technical field, and provides a gene combination used for guiding individual treatment of platinum medicines. The gene combination comprises GSTP1, MRP2, XRCC1-Exon6, XRCC1-Exon10, ERCC1 and ERCC2. By detecting the mutation condition of gene sites of GSTP1, MRP2, XRCC1-Exon6, XRCC1-Exon10, ERCC1 and ERCC2 in tumor tissues of patients, the gene combination carries out a medicine sensitive guidance to the patients treated by platinum medicines for maximizing the curative effect and minimizing the side-effect.

Description

technical field [0001] The invention belongs to the field of gene technology and relates to a group of gene combinations used to guide individualized treatment of platinum drugs. Background technique [0002] Platinum chemotherapy drugs (cisplatin, carboplatin, oxaliplatin, etc.) are one of the most commonly used drugs in the field of tumor chemotherapy. Formation of platinum-DNA adducts triggers a series of intracellular events that ultimately lead to tumor cell death. However, the poor sensitivity of some tumor cells to platinum-based drugs is the main reason for the failure of chemotherapy and the progression of the disease. Therefore, before using platinum-based drugs, clinicians urgently need a piece of prospective guiding information on efficacy, drug resistance, and prognosis. [0003] GSTP1 belongs to the family of multifunctional phase II metabolic enzymes, which can catalyze the combination of glutathione and various toxic compounds (including platinum-based prep...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 楼敬伟
Owner SHANGHAI BIOTECAN PHARMA
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