Method for detecting PTEN (Phosphatase and tensin homolog) gene and PI3K/AKT protein and application of method in cancer treatment

A technology in the gene and sequence table, applied in the field of detection of PTEN gene and PI3K/AKT protein, can solve the problem of low objective response rate, and achieve the effect of preventing the missed treatment opportunity and economic loss, and the detection result is accurate.

Inactive Publication Date: 2015-04-01
BEIJING QINLAN BIOTECH
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Problems solved by technology

Trastuzumab is used to treat metastatic breast cancer patients with HER2 overexpression, and has achieved good therapeutic effect, but the objective response rate of its single treatment is not high, and the median survival period of 9 months is only 12%-34 %, and most patients develop resistance within one year of using the drug

Method used

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  • Method for detecting PTEN (Phosphatase and tensin homolog) gene and PI3K/AKT protein and application of method in cancer treatment
  • Method for detecting PTEN (Phosphatase and tensin homolog) gene and PI3K/AKT protein and application of method in cancer treatment
  • Method for detecting PTEN (Phosphatase and tensin homolog) gene and PI3K/AKT protein and application of method in cancer treatment

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Embodiment 1

[0052] A method for detecting PTEN gene, said detection method comprising the following steps:

[0053] DNA sequencing operations

[0054] 1) Total DNA extraction: Take 1cm of fresh breast tissue (including cancer tissue and paracancerous tissue) from patients with breast cancer (female A and woman B) 3 ×1cm 3 ×1cm 3 size, washed with normal saline for 3 times, and cut the tissue into pieces with scissors, added DNA extraction buffer (0.33M NaAc, 25mM EDTA, pH6.4), 10% SDS and proteinase K (10mg / ml), 55°C Water bath for 3 hours, and mix well by inverting during the water bath; after the water bath, add 500 μl of isopropanol to the tissue, gently invert and mix for 10 minutes, then centrifuge at 12000 rpm for 15 minutes, discard the supernatant, the precipitate is DNA precipitation, and DNA precipitation After air-drying, 100 μl of TE buffer was added to the DNA pellet to dissolve the DNA pellet, and the concentration of the DNA was measured in a Nano Drop (DNA UV spectropho...

Embodiment 2

[0074] A method for detecting PI3K / AKT protein, said detection method comprising the following steps:

[0075] Western Blotting operation

[0076] 1) Take fresh breast tissue from patients with breast cancer (female A and female B), place the tissue on ice, and separate the tissue with a sterilized pre-cooling tool to obtain tissue blocks;

[0077] 2) Put the tissue block into a round-bottomed Eppendorf tube, add liquid nitrogen to the tube to freeze the tissue block, place the Eppendorf tube on ice, and grind it evenly to obtain tissue grinds;

[0078] 3) Add lysis buffer to the tissue ground material, the amount of the lysis solution added is 60 μl / mg, homogenize in ice bath, and then shake the Eppendorf tube at 4°C for 2 hours to obtain a homogeneous tissue Slurry, the protein concentration in the tissue homogenate reaches at least 0.1 mg / ml; the lysis buffer is purchased from Beyontian Company;

[0079] 4) Centrifuge the tissue homogenate at 4°C for 20 minutes at a speed...

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Abstract

The invention provides a method for detecting a PTEN gene and a PI3K (Phosphatidyl Inositol 3-kinase) / AKT (Protein Kinase B) protein and an application of the method in breast cancer individuation treatment. The method is characterized in that a DNA (deoxyribonucleic Acid) sequencing method is used for detecting the PTEN in pathological tissues of breast cancer patients, a WesternBlot technology is used for detecting the expression condition of the PI3K / AKT protein, and detection results are accurate and reliable. The detection method provided by the invention can be used for accurately judging the tolerance of patients to trastuzumab in combination with the sensitivity of the breast cancer patients to the trastuzumab, providing the basis for the diagnosis, treatment and evaluation of the breast cancer patients and preventing the trastuzumab (Herceptin) from being blindly used to cause the missing of treatment opportunity and the economic loss for the patients.

Description

technical field [0001] The invention belongs to the field of nucleic acid and protein detection methods, in particular to a method for detecting PTEN gene and PI3K / AKT protein and its application. Background technique [0002] Breast cancer is currently the highest incidence rate among women worldwide and is the leading cause of cancer death in women. With the aging of the global population and the continuous growth of the world's total population, as well as the backwardness of medical facilities in developing countries and regions, the incidence of cancer has risen sharply. According to the "Global Cancer Statistics" global tumor statistics report published online by A Cancer Journal for Clinicians in 2011, about 12.7 million cancer cases and 7.6 million cancer deaths were assessed, of which about 56% of cancer cases and 64% of Cancer deaths were from developing countries or regions. Breast cancer has a high incidence in women and is the leading cause of cancer death in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N33/68
Inventor 贾浩路志越丛茜郭晗江丽娜胡博廖博
Owner BEIJING QINLAN BIOTECH
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