Method for constructing non-small-cell lung cancer gefitinib drug-resistance PDX model

A technology for non-small cell lung cancer and non-small cell cancer, applied in the direction of medical raw materials derived from mammals, can solve the problem of difficult to obtain drug-resistant tissue PDX model construction, and achieve good promotion and application value

Inactive Publication Date: 2016-02-03
ZHENGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Deficiencies of the first construction method: 1. Gefitinib-resistant non-small cell lung cancer patients are usually in the advanced stage of cancer and have no chance to undergo surgery. Usually, re-diagnosis is performed by puncture, and the amount of tissue obtained by puncture is small, so it is difficult Obtain enough drug-resistant tissues for PDX model construction
2 Gefitinib resistance is the occurrence of drug resistance after taking gefitinib for a long time. Before the occurrence of gefitinib resistance, patients cannot be punctured in stages to obtain the tissue in the process of gefitinib resistance.

Method used

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  • Method for constructing non-small-cell lung cancer gefitinib drug-resistance PDX model
  • Method for constructing non-small-cell lung cancer gefitinib drug-resistance PDX model
  • Method for constructing non-small-cell lung cancer gefitinib drug-resistance PDX model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] This example mainly introduces the construction process of the PDX model of non-small cell lung cancer (NSCLC).

[0048] 1. Obtain tissue samples from patients with non-small cell lung cancer

[0049] The obtained non-small cell carcinoma tumor tissue samples come from fresh tissue samples after surgery. Generally speaking, when further selecting tumor samples, marginal tumor tissues should be taken as much as possible to avoid necrotic tissue in the collected tissues.

[0050] According to needs, the size of the finally selected tumor tissue sample should be 1 cm×1 cm. After photographing and weighing the selected tumor tissue samples and making records, for the convenience of subsequent detection and experiments, the obtained tumor tissue samples can be divided into three parts: one part is fixed with 10% formalin, and paraffin wax is made Specimens were used for pathological examination and immunohistochemical analysis; a part was refrigerated at -80°C for DNA / RNA...

Embodiment 2

[0068] This example mainly introduces the evaluation of the non-small cell lung cancer PDX model constructed in Example 1 and the process of screening the gefitinib-sensitive non-small cell lung cancer PDX model.

[0069] The evaluation of the non-small cell lung cancer PDX model constructed in Example 1 mainly includes two aspects of histopathological evaluation and EGFR gene molecular evaluation, as follows.

[0070] Tumor growth curve should be drawn first in histopathological evaluation ,Specifically:

[0071] As in the calculation method of the tumor volume described in Example 1, the tumor growth volume is measured and calculated once a week, and the tumor growth curve is drawn accordingly; at the same time, the critical volume value of the tumor growth (that is, the tumor volume value at the time of passage) should be recorded and determined. ;

[0072] It should be emphasized that the critical volume of tumor growth in mice in the specific non-small cell lung cancer...

Embodiment 3

[0087] On the basis of the above-mentioned Examples 1 and 2, the inventor screened and obtained a tumor growth critical value of 500mm 3 1. The PDX model of non-small cell lung cancer with EGFR mutation type L858R. This example mainly introduces the process of inducing gefitinib on this PDX model to obtain a gefitinib-resistant PDX model of non-small cell lung cancer.

[0088] For the non-small cell lung cancer PDX model obtained through screening, the third-generation xenograft mice were used to induce and construct a xenograft tumor model that acquired resistance to gefitinib; the specific process was as follows:

[0089] Mice are divided into two groups, and one group is control group, and another group is experimental group, and positive control group is administrated with Gefitinib solution, every day 100mg / kg, and tumor volume can be obviously dwindled (positive control group) after continuous intragastric administration for a week. The significance that group is set up:...

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Abstract

The invention belongs to the technical field of lung cancer gefitinib drug-resistance model construction, and particularly relates to a method for constructing a non-small-cell lung cancer gefitinib drug-resistance PDX model. The method includes the steps of constructing a non-small-cell lung cancer PDX model, screening a gefitinib-sensitive non-small-cell lung cancer PDX model and induction constructing of the non-small-cell lung cancer gefitinib drug-resistance PDX model. The model can be used as a substitution of the PDX model which is directly established by non-small-cell lung cancer gefitinib drug-resistance tumor issue. The good research basis is provided for studying gefitinib drug-resistance mechanisms, screening target spots and small molecule drugs in the gefitinib drug-resistance process and hence delaying or interfering of the gefitinib drug-resistance process, and good application and popularization value is achieved.

Description

technical field [0001] The invention belongs to the technical field of constructing a gefitinib-resistant model of lung cancer, and specifically relates to a patent application for a method for constructing a gefitinib-resistant PDX model of non-small cell lung cancer. Background technique [0002] Lung cancer ranks first in cancer mortality worldwide, and non-small-cell lung cancer (NSCLC) accounts for 80% of lung cancer. The 5-year survival rate of non-small cell lung cancer is less than 15%. The main reason is that about 70% of patients with non-small cell lung cancer are already at an advanced stage when they are first diagnosed, and most of them have distant metastasis. Lung cancer is one of the first types of cancer to use targeted therapy in the clinic. In the past decade, small-molecule compound tyrosine kinase inhibitors have been widely used in the clinical targeted therapy of NSCLC patients, and the target is epidermal growth factor receptor (Epidermal Growth Fac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/13A61K31/5377
Inventor 刘康栋江亚南赵继敏张翼董子钢董子明吴琼尹学善王艳红
Owner ZHENGZHOU UNIV
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