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Oral squarmous cell carcinomas pathogenic gene BPIFB2 and application thereof

An oral cancer and gene technology, applied in the field of bioengineering, can solve the problems of unknown invasion and metastasis mechanism, lack of early prevention and prognosis judgment means, unclear pathogenesis of oral cancer, etc., and achieve the effect of ensuring consistency and convenient clinical use

Inactive Publication Date: 2014-09-24
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The main reasons for this situation are: the pathogenesis of oral cancer is unclear, the mechanism of invasion and metastasis is unknown, and there is a lack of effective means of early prevention and prognosis.

Method used

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  • Oral squarmous cell carcinomas pathogenic gene BPIFB2 and application thereof
  • Oral squarmous cell carcinomas pathogenic gene BPIFB2 and application thereof
  • Oral squarmous cell carcinomas pathogenic gene BPIFB2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Collection of research samples and extraction of RNA

[0026] Eight cases of oral cancer and normal fresh tissues were collected, grouped and numbered: 3 cases of normal tissue (numbered N1, N2, N3), 3 cases of paracancerous tissue (numbered P1, P2, P3) and 2 cases of Cancer tissue blocks (C1 and C2).

[0027] The three groups of samples were subjected to RNA extraction, followed by agarose gel electrophoresis after RNA extraction. From the results of electrophoresis, the quality of the extracted RNA samples was preliminarily considered to be qualified and could be used for further transcriptome analysis. Further, the extraction of RNA samples was detected by NanoDrop1000 spectrophotometer, and the results are shown in Table 1. Each sample volume was 30 μl. Sample requirements for RNA-seq sequencing: OD260 / OD280 is 1.8-2.2. RIN (RNA integrity number) refers to the RNA integrity index, which is an important indicator of RNA quality. It can be seen from Tab...

Embodiment 2

[0030] Example 2 High-throughput transcriptome sequencing and analysis

[0031] Sequencing platform: Illumina's HiSeq2500 high-throughput sequencing platform

[0032] Transcriptome sequencing and matching:

[0033]The three groups of cancerous, paracancerous and normal tissue samples we analyzed came from 8 patients with oral cancer. These three sets of samples were subjected to high-throughput cDNA sequencing. From these three tissues we obtained 4.85×107, 4.91×107, 4.46×107 read pairs, respectively. We used TopHat to match reads to the UCSC reference human genome (hg19), the unique matched read pairs ranged from 4.04×107 to 4.49×107, and the proportion of reads matching to the Ensembl reference gene was between 81% and 89% between. The average coverage of our sequencing depth is about 130 times that of the human genome (according to the total length of the exon region uniquely annotated in the Ensembl database, about 1.13×108), and only 1% of the reads are mapped to rRNA...

Embodiment 3

[0040] Example 3 BPIFB2 gene expression in oral cancer tissue and adjacent tissue

[0041] 1. Materials and methods

[0042] 1. Materials

[0043] Oral cancer patients were hospitalized from 2005 to 2010, and tumor tissues and paracancerous tissues were collected from 50 patients with oral cancer.

[0044] 2. Method

[0045] 2.1 Extraction of total RNA from oral cancer tissues and paracancerous tissues

[0046] Extract RNA from oral cancer tissue and paracancerous tissue according to the instructions of Kangwei Century Ultrapure RNA Extraction Kit (Ultrapure RNA Kit (DNase I), Cat. No. CW0597), prove the integrity of RNA by gel electrophoresis, and measure RNA with a nucleic acid protein analyzer concentration and purity.

[0047] Use the Ultrapure RNA Extraction Kit (Cat. No. CW0597) to extract, and the main steps are as follows:

[0048] 1. Sample processing, 30-50mg tissue is fully ground in liquid nitrogen and then added 1ml Buffer RLT, or 1ml Buffer RLT is added to t...

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Abstract

The invention relates to an oral squarmous cell carcinomas pathogenic gene BPIFB2 and application of the oral squarmous cell carcinomas pathogenic gene BPIFB2. RNA is extracted from oral squarmous tissue, tissue adjacent to the oral squarmous cell carcinomas and normal tissue, deep sequencing and analyzing are performed on a transcriptome, the oral squarmous cell carcinomas relevant gene BPIFB2 is obtained through screening, an RT-PCR method is adopted for analyzing the expression conditions, on the carcinomas tissue and the tissue adjacent to the carcinomas of a patient with the oral squarmous cell carcinomas, of the screened BPIFB2, and the result shows that the obtained BPIFB2 through screening has good relevance to the oral squarmous cell carcinomas and can be used for preparing an oral squarmous cell carcinomas auxiliary diagnosis preparation or a pregnosis preparation. The invention further discloses a fluorogenic quantitative PCR kit and a using method of the fluorogenic quantitative PCR kit. The kit can detect the expression level of the BPIFB2 in oral squarmous cell carcinomas tissue sensitively, rapidly, quantitatively, accurately and stably and has good application prospects.

Description

technical field [0001] The present invention relates to the technical field of bioengineering, in particular, the present invention relates to the screening of the oral cancer pathogenic gene BPIFB2 through high-throughput transcriptome sequencing analysis and the application of the pathogenic gene in the preparation of auxiliary diagnostic or prognostic preparations for oral cancer. Background technique [0002] Oral Squarmous Cell Carcinomas (OSCC) is one of the common tumors in the human body, especially in developing countries such as India, Sri Lanka, and Brazil, where its incidence can reach 25% of all tumors. In recent years, the incidence of oral cancer has a clear upward trend in developed countries such as Europe and the United States, especially the age of onset tends to be younger. Oral squamous cell carcinoma not only has a high incidence rate, but also has a high degree of malignancy; it often causes dysfunction of speech, chewing, swallowing, and facial featur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06
CPCC12Q1/6851C12Q1/6886C12Q2600/118C12Q2600/158C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 杨承刚李红伟常鹏孙锦云
Owner BEIJING MEDINTELL BIOMED CO LTD
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