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Application of TEX19 gene in diagnosis and treatment of biliary duct cancer

A cholangiocarcinoma and gene technology, which can be used in gene therapy, medical preparations containing active ingredients, and antibody medical components, etc., can solve the problems of limited diagnostic methods, low surgical resection and radical cure rates, and lack of diagnostic methods for human cholangiocarcinoma.

Inactive Publication Date: 2015-09-09
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the diagnostic methods for human cholangiocarcinoma are still limited. Imaging techniques such as B-ultrasound, CT, and MRI are still the main diagnostic methods for human cholangiocarcinoma. It is an advanced tumor with low surgical resection rate and radical cure rate. At present, there is still a lack of a specific and sensitive diagnostic method for human cholangiocarcinoma

Method used

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  • Application of TEX19 gene in diagnosis and treatment of biliary duct cancer
  • Application of TEX19 gene in diagnosis and treatment of biliary duct cancer
  • Application of TEX19 gene in diagnosis and treatment of biliary duct cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Screening for gene markers associated with cholangiocarcinoma

[0054] 1.1 Sample collection

[0055] Eight samples of normal bile duct tissue and cholangiocarcinoma tissue were collected. The above samples are surgical resection specimens of patients with cholangiocarcinoma, and all the above samples were obtained with the consent of the organizational ethics committee.

[0056] 1.2 RNA sample preparation and quality analysis

[0057] 1.2.1 Preparation of RNA samples

[0058] Total RNA in advance using QIAGEN Tissue RNA Extraction Kit. Specific steps are as follows:

[0059] 1) In a clean area with less RNase interference, use a mortar containing an appropriate amount of liquid nitrogen to weigh about 20 mg of the isolated liver cancer tissue sample, and grind it to powder with a pestle;

[0060] 2) Transfer the sample to a RNase-free 2mL centrifuge tube;

[0061] 3) Add 300 μl Lysis solution, place in a homogenizer, and grind thoroughly for 1-5 minutes...

Embodiment 2

[0098] Example 2 RT-PCR sequencing to verify the differential expression of TEX19 gene

[0099] 1. According to the detection results of high-throughput sequencing, the TEX19 gene was selected for large-sample RT-PCR verification. According to the sample collection method in Example 1, 80 cases of cholangiocarcinoma tissue and 80 cases of normal bile duct tissue were selected.

[0100] 2. The RNA extraction steps are the same as in Example 1.

[0101] 3. Reverse transcription: use the reverse transcription kit of TAKARA company to operate. Specific steps are as follows:

[0102] (1) Take 2 μg of total RNA for reverse transcription, add 2 μl of Oligo(dT), and mix well. After 5 minutes in water bath at 70°C, immediately ice bath for 2-3 minutes.

[0103] (2) Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), 40 U / μl of RNasin, 200 U / μl of M-MLV, and make up to the expected volume with nuclease-free water.

[0104] (3...

Embodiment 3

[0125] Example 3 Inhibition of TEX19 gene expression

[0126] 1. Cell culture: human cholangiocarcinoma cell line QBC939, with DMEM (high glucose) medium containing 10% calf serum at 37°C, 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. The medium was changed once every 2-3 days, and 0.25% trypsin was used for routine digestion and passage.

[0127] 2. siRNA design

[0128] siRNA sequence against TEX19:

[0129] siRNA1-TEX19:

[0130] The sense strand is 5'-UUGAACUUCUGGAAAUGUCUU-3' (SEQ ID NO.7);

[0131] The antisense strand is 5'-GACAUUUCCAGAAGUUCAAGC-3' (SEQ ID NO.8),

[0132] siRNA2-TEX19:

[0133] The sense strand is 5'-UGUUGAAGCUGAUACAUCCAG-3' (SEQ ID NO.9);

[0134] The antisense strand is 5'-GGAUGUAUCAGCUUCAACAUG-3'(SEQ ID NO.10),

[0135] siRNA3-TEX19:

[0136] The sense strand is 5'-UAACGAUGUUUCUAGAAUGCU-3' (SEQ ID NO.11);

[0137] The antisense strand is 5'-CAUUCUAGAAACAUCGUUAAC-3' (SEQ ID NO.12,

[0138] Negative control siRNA seque...

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Abstract

The invention discloses a TEX19 gene and an expression product of the TEX19 gene serving as a molecular marker in early diagnosis of biliary duct cancer. In the invention, RNA-sep is used for screening, large-sample RT-PCR is used for verification, and TEX19 gene expression abnormity is related to generation and development of biliary duct cancer. According to the search achievements, a drug capable of inhibiting TEX19 gene expression in biliary duct tissue or inhibiting TEX19 gene from expressing product functions can be searched, thus realizing prevention and treatment of biliary duct cancer on clinic.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of human TEX19 gene in the diagnosis and treatment of cholangiocarcinoma. Background technique [0002] Human cholangiocarcinoma is the second most common hepatobiliary malignancy after liver cancer. At present, surgical resection or liver transplantation is the only effective treatment that may cure human cholangiocarcinoma. However, due to the insidious onset and strong invasiveness of human cholangiocarcinoma, most patients have lymphatic or distant metastases when they see a doctor, resulting in a low surgical cure rate. , so the overall survival prognosis is very poor, and its 5-year survival rate is only 20%-40%. Early prevention, early detection, and early treatment are the key to improving the curative rate and 5-year survival rate of human cholangiocarcinoma. [0003] At present, the diagnostic methods for human cholangiocarcinoma are still limited, and ima...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K48/00A61K31/713A61K39/00A61K45/00A61P35/00
CPCA61K39/0005A61K45/00G01N33/57407G01N33/57484G01N33/6893A61K31/713C12Q1/6886C12Q2600/158
Inventor 杨承刚果春青宋宏涛
Owner BEIJING MEDINTELL BIOMED CO LTD
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