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Copy number amplification of FRS2 gene, applications of copy number amplification, and specific primer pairs for detecting copy number amplification

A technology of gene copy number and copy number, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., which can solve the problems of lack of risk grading method, inability to judge chemotherapy sensitivity and recurrence tendency, etc. , to achieve the effect of large blood vessel density

Active Publication Date: 2019-01-22
SHENZHEN LUOHU PEOPLELS HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The results of pathological diagnosis of bladder cancer play an important role in the selection of treatment options, but when judging the malignancy of tumors, there is a lack of objective and accurate risk classification methods, and it is impossible to judge chemotherapy sensitivity and recurrence tendency before treatment

Method used

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  • Copy number amplification of FRS2 gene, applications of copy number amplification, and specific primer pairs for detecting copy number amplification
  • Copy number amplification of FRS2 gene, applications of copy number amplification, and specific primer pairs for detecting copy number amplification
  • Copy number amplification of FRS2 gene, applications of copy number amplification, and specific primer pairs for detecting copy number amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Sample collection and whole genome sequencing analysis:

[0038] In accordance with medical ethics and with the patient's consent, 65 pairs of paired bladder tumor tissue and peripheral blood samples were obtained from newly diagnosed individuals with bladder cancer in the Institute of Urology, Shenzhen University for whole genome sequencing analysis of 65 cases of bladder cancer The clinical information of the patients is shown in Table 1.

[0039] Table 1 Clinical information table of 65 bladder cancer patients analyzed by whole genome sequencing

[0040]

[0041]

[0042]

[0043] Genomic DNA was extracted from 65 bladder cancer patients shown in Table 1 for whole-genome sequencing. The specific method is as follows:

[0044]1. Raw data was filtered using SOAPnuke (v.1.5) to remove adapter sequences and low-quality reads. The high-quality reads were aligned with the human reference genome (hg19, NCBI GRCh37) by Burrows-Wheeler Aligner (BWA) a...

Embodiment 2

[0049] Example 2: Validation of FRS2 copy number amplification:

[0050] In the case of conforming to medical ethics, 196 samples of bladder cancer who had not undergone radiotherapy and chemotherapy were screened from the paraffin sections of bladder cancer preserved in the sample bank of the Institute of Urology, Shenzhen University. The information of the 196 samples is shown in Table 2.

[0051] Table 2 The clinical information table of 196 patients with bladder cancer targeted verification

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058] In order to further verify the copy amplification of FRS2 gene in bladder cancer, specific primers for FRS2 were designed, GAPDH gene was used as internal reference, and blood cells and urine exfoliated cells of healthy people were used as control. Targeted validation was performed in newly enrolled 196 bladder cancer samples. The specific steps are:

[0059] For the 196 samples shown in Table 2, use th...

Embodiment 3

[0074] Example 3: Effect of FRS2 copy number amplification on FRS2 expression and microangiogenesis in tumor tissue:

[0075] In order to further explore the effect of copy number amplification of the FRS2 gene on the tumor in bladder cancer, the expression of the protein encoded by the FRS2 gene in tumor tissue was analyzed by immunohistochemical method, and based on the biological function of FRS2 to promote angiogenesis , analyzed the effect of FRS2 copy number amplification on vessel density in tumor tissues.

[0076] In 196 cases of bladder cancer tumor samples with FRS2 copy number amplification, the expression of FRS2 protein in high copy and low copy samples of FRS2 gene and the formation of microvessels in tumor tissue were analyzed by immunohistochemical method. Specific steps are as follows:

[0077] The number of microvessels was assessed by immunohistochemistry for CD31. Using the specific antibodies anti-FRS2 (Abcam, catalog number ab150058) and anti-CD31 (Ab...

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Abstract

The invention provides copy number amplification of the FRS2 gene, applications of the copy number amplification, and specific primer pairs for detecting the copy number amplification, and belongs tothe field of biotechnologies. The number of times for copy number amplification is 3 or more. The density of the new blood vessels in the bladder cancer sample after the copy number amplification is high. In the bladder cancer cell line cultured in vitro, specific siRNA is utilized for interfering the expression of FRS2, so that the capacity of recruiting the vascular endothelial cells of the bladder cancer cells and the capacity of inducing the vascular endothelial cells to form tubules of the bladder cancer cells are inhibited. The invention further provides the applications of the copy number amplification of the FRS2 gene in preparing kits for detecting bladder cancer. The technical scheme has the advantages that the copy number amplification can serve as one of biological detection indexes of a molecular probe or a targeting primer for developing the bladder cancer screening kit.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the copy number amplification of the FRS2 gene and its application, and a pair of specific primers for detecting the copy number amplification. Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system. The pathological types of bladder cancer in WHO's "Pathology and Genetics of Tumors of the Urinary System and Male Genital Organs" in 2004 include bladder urothelial carcinoma, bladder squamous cell carcinoma, bladder adenocarcinoma, and other rare ones include bladder clear cell carcinoma, Small cell carcinoma of the bladder, bladder carcinoid. The most common of these is urothelial carcinoma of the bladder, accounting for more than 90% of the total number of bladder cancer patients. Bladder cancer is usually referred to as urothelial carcinoma of the bladder, which was previously called transitional cell carcinoma of the bladd...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158
Inventor 吴松欧铜崔香蕊
Owner SHENZHEN LUOHU PEOPLELS HOSPITAL
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