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Novel application of BPIFB1 to preparing tongue squamous cell carcinoma diagnosis and treatment preparations

A technology of tongue squamous cell carcinoma and reagents, applied in the treatment, BPIFB1 gene in the field of diagnosis of tongue squamous cell carcinoma, can solve the problems of inability to further expand microquantification, inability to form scale, and expensive sequencing methods

Inactive Publication Date: 2016-08-24
XIANGYA STOMATOLOGICAL HOSPITAL CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the new generation technology, the traditional sequencing technology Sanger sequencing method has certain limitations. In addition to the high cost of the sequencing method, it also relies heavily on electrophoretic separation technology, which cannot be further expanded and microquantified, so it cannot be scaled up.

Method used

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  • Novel application of BPIFB1 to preparing tongue squamous cell carcinoma diagnosis and treatment preparations
  • Novel application of BPIFB1 to preparing tongue squamous cell carcinoma diagnosis and treatment preparations
  • Novel application of BPIFB1 to preparing tongue squamous cell carcinoma diagnosis and treatment preparations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Differences in the expression of BPIFB1 gene between tongue squamous cell carcinoma tissue and adjacent tissue

[0059] 1. Experimental materials:

[0060] The 20 cases of tongue squamous cell carcinoma and paracancerous tissue samples were taken from patients who underwent radical resection of tongue squamous cell carcinoma. All tissue samples were confirmed by histopathology, and a consent form was signed for voluntary donation for use in this experiment. The specimens were collected within 30 minutes of the surgical procedure, and then collected under the guidance of pathologists. The size of each sample is about 1.0cm*l.0cm*l.0cm. The cancerous tissue was the central area of ​​the tumor when the material was collected; the corresponding paracancerous tissue was the tissue 1-2cm away from the edge of the cancerous tissue. All specimens were placed in 1.5ml EP tubes and stored in liquid nitrogen tanks. All specimens were collected from patients who had no...

Embodiment 2

[0150] Example 2 Large sample verification differentially expressed gene BPIFB1 gene

[0151] 1. Experimental materials

[0152] According to the method of Example 1, 50 tongue squamous cell carcinoma tissues and their corresponding paracancerous tissues were collected.

[0153] 2. RNA extraction

[0154] With embodiment 1.

[0155] 3. Reverse transcription

[0156] The reverse transcription of RNA was carried out using the reverse transcription kit of TAKARA company.

[0157] 4. QPCR

[0158] (1) Primer design

[0159] QPCR amplification primers were designed according to the coding sequences of BPIFB1 gene and GAPDH gene in Genbank, and synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. The specific primer sequences are as follows:

[0160] BPIFB1 gene:

[0161] The forward primer is 5'-ACCAACTTATACTCAACTT-3' (SEQ ID NO.3);

[0162] The reverse primer is 5'-GATCTCAGTGATGATGTT-3' (SEQ ID NO.4),

[0163] GAPDH gene:

[0164] The forward primer ...

Embodiment 3

[0175] Example 3 BPIFB1 Gene Expression Plasmid Construction

[0176] 1. Construction of BPIFB1 gene expression vector

[0177] Amplification primers were designed according to the coding sequence of the BPIFB1 gene (as shown in SEQ ID NO.1). Amplify the coding sequence of the full-length BPIFB1 gene from the cDNA library of adult fetal brain (clontech company, article number: 638831), insert the above cDNA sequence into the eukaryotic cell expression vector pcDNA3.1, and connect the obtained recombinant vector pcDNA3.1 -BPIFB1 was used in subsequent experiments.

[0178] 2. Culture and transfection of tongue squamous cell carcinoma cells

[0179] Cell culture: Take the tongue squamous cell line Tca8113 and culture it in DMEM containing 10% calf serum at 37°C and 5% CO 2 cultured in a cell culture incubator.

[0180] Tongue squamous cell carcinoma cells by 1×10 4 Inoculate / well into 24-well cell culture plates at 37°C, 5% CO 2 The cells were cultured in the incubator for...

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Abstract

The invention discloses a gene BPIFB1 and an expression product thereof. The gene BPIFB1 is a molecular marker for tongue squamous cell carcinoma early diagnosis. The gene BPIFB1 and the expression product have the advantages that as proved by QPCR (quantitative polymerase chain reaction), the gene BPIFB1 can be differently expressed in tongue squamous cell carcinoma tissues and para-carcinoma tissues and can be used as an indicator for tongue squamous cell carcinoma early diagnosis; the gene BPIFB1 and the expression product can be used as targets for treating tongue squamous cell carcinoma and can be used for guiding research and development of new medicines.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of BPIFB1 gene in the diagnosis and treatment of squamous cell carcinoma of the tongue. Background technique [0002] Oral squamous cell carcinoma is the most common malignant tumor of the head and neck, accounting for about 3% of systemic tumors and 90% of oral tumors, and tongue cancer ranks first in the incidence of oral squamous cell carcinoma. The International Agency for Research on Cancer found that there were 300,373 new cases and 145,353 deaths from oral cancer in 2012. In my country, there were 39,450 new cases and 16,933 deaths in 2011. Male morbidity and mortality were higher than For women, urban is higher than rural. The incidence of tongue squamous cell carcinoma in my country is increasing year by year. Because most patients have no obvious clinical symptoms in the early stage, they are often not paid attention to. Once clinical symptoms appear, it is already...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/57484G01N33/68G01N2800/18
Inventor 刘欧胜王月红卢燕勤易桥周典黄宏宇
Owner XIANGYA STOMATOLOGICAL HOSPITAL CENT SOUTH UNIV
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