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Applications of TFF2 gene as intracranial aneurysm diagnosis and treatment marker

An intracranial aneurysm and gene technology, which can be used in the determination/examination of microorganisms, medical preparations containing active ingredients, disease diagnosis, etc., and can solve the problems of high false negative rate and high cost.

Inactive Publication Date: 2016-03-16
SICHUAN PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

All three methods have the disadvantages of high cost and high false negative rate. At the same time, DSA, which is the gold standard, is still an invasive test, which is difficult to be accepted by the general population. Early diagnosis and screening of patients, and corresponding effective treatment, ultimately improve the overall prognosis of patients

Method used

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  • Applications of TFF2 gene as intracranial aneurysm diagnosis and treatment marker
  • Applications of TFF2 gene as intracranial aneurysm diagnosis and treatment marker
  • Applications of TFF2 gene as intracranial aneurysm diagnosis and treatment marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Expression difference of TFF2 gene in intracranial aneurysm tissue and normal control tissue

[0059] 1. Sample collection

[0060] Collected 40 patients with intracranial aneurysms, surgically clipped the neck of the aneurysm, removed the aneurysm specimens, removed the adhesion tissue and wall thrombus, and immediately put them into liquid nitrogen for storage. The superficial temporal arteries of 30 patients with traumatic brain injury were taken and put into liquid nitrogen for preservation as normal control tissues. Aged 40-60 years old, no special treatment before operation. The samples were collected with the consent of the patients and their families.

[0061] 2. Extraction of tissue total RNA

[0062] The total RNA in the tissue was extracted using the Trizol one-step method, and the purity of the RNA solution was determined by reading the absorbance values ​​at 260nm and 280nm (A) by NanodropND-1000. The integrity of RNA was detected by electroph...

Embodiment 2

[0073] Embodiment 2 interferes with the expression of TFF2 gene

[0074] 1. siRNA design and synthesis

[0075] siRNA sequence against TFF2:

[0076] siRNA1-TFF2:

[0077] The sense strand is 5'-AGAGAUAACCUUUCUUCUCUG-3' (SEQ ID NO.3);

[0078] The antisense strand is 5'-GAGAAGAAAGGUUAUCUCUGC-3' (SEQ ID NO.4),

[0079] siRNA2-TFF2:

[0080] The sense strand is 5'-ACUUCAAAGAUGAAGUUGGAG-3' (SEQ ID NO.5);

[0081] The antisense strand is 5'-CCAACUUCAUCUUUGAAGUGC-3' (SEQ ID NO.6),

[0082] siRNA3-TFF2:

[0083] The sense strand is 5'-AUUAUAUGUUAAACCAUUGAA-3' (SEQ ID NO.7);

[0084] The antisense strand is 5'-CAAUGGUUUAACAUAUAAUUU-3'(SEQ ID NO.8)

[0085] Negative control siRNA sequence (siRNA-NC):

[0086] The sense strand is 5'-CGUACGCGGAAUACUUCGA-3' (SEQ ID NO.9);

[0087] The antisense strand is 5'-UCGAAGUAUUCCGCGUACG-3' (SEQ ID NO.10).

[0088] 2. Cell culture

[0089] Take the vascular smooth muscle cells HA-VSMC and culture them with high glucose 1640 with a conce...

Embodiment 3

[0102] Example 3 Effect of TFF2 gene on apoptosis of vascular smooth muscle cells

[0103] Use AnnexinV-FITC / PI double staining method to detect cell apoptosis by flow cytometry, the steps are as follows:

[0104] 1. Cell transfection: The vascular smooth muscle cells were transfected with siRNA1-TFF2 and siRNA-NC according to the method in Example 2.

[0105] 2. After 48 hours of cell transfection, the cells were harvested, and the cells were processed according to the operating instructions of the AnnexinV-FITC / PI Double Staining Kit (purchased from Invitrogen).

[0106] 3. The cells treated in step 2 are detected by flow cytometry.

[0107] 4. Statistical methods

[0108] Statistical cell survival rate, cell survival rate = 1-apoptosis rate, the experiment is done by repeating 3 times, the result data is expressed in the form of mean ± standard deviation, using SPSS13.0 statistical software to carry out For statistical analysis, the difference between the two was tested ...

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Abstract

The invention discloses applications of TFF2 gene and expression products of TFF2 gene as intracranial aneurysm diagnosis and treatment molecular markers. It can be determined whether a patient is suffered from intracranial aneurysm or the patient possesses the risk of suffering from intracranial aneurysm based on the content of TFF2 gene and the expression products of TFF2 gene in the arterial tissues of the patient. It is found via study on apoptosis conditions of in-vitro cultured vascular smooth muscle cells that interference of TFF2 gene expression is capable of promoting vascular smooth muscle cell apoptosis, so that it is shown by the above results that TFF2 gene and the expression products of TFF2 gene are potential drug targets in treatment of intracranial aneurysm.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of human TFF2 gene in the diagnosis and treatment of intracranial aneurysms. Background technique [0002] Intracranial aneurysm (IA) is a tumor-like lesion formed by the outward protrusion of the intracranial artery wall, and is one of the more common cerebrovascular diseases. The incidence rate of intracranial aneurysm in the crowd is 29%-59%. Once it ruptures and bleeds, the rate of disability and death is very high. A large number of case studies have shown that both microsurgery and endovascular embolization can achieve satisfactory results. If effective prevention and treatment are carried out before IA rupture, the overall prognosis of patients will be greatly improved. Currently, the diagnosis and screening of intracranial aneurysms mainly rely on cerebral angiography (DSA), magnetic resonance angiography (MRA) and CT angiography (CTA). All three methods ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68A61K45/00A61P35/00
CPCA61K45/00C12Q1/6883C12Q2600/158G01N33/6893G01N2800/329
Inventor 李志立黄光富陈隆益谭海斌王振宇石毅刘泠殷成王奇
Owner SICHUAN PROVINCIAL PEOPLES HOSPITAL
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