Application of LCE3E to diagnosis and treatment of oral cancer

A technology of oral cancer and preparations, applied in the field of bioengineering, can solve problems such as unclear development mechanism, achieve clear results, improve work efficiency, and have good stability

Active Publication Date: 2016-03-09
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The occurrence and development of oral squamous cell carcinoma involves the changes of many different genes. However, its exact mechanism of occurrence and development is s

Method used

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  • Application of LCE3E to diagnosis and treatment of oral cancer
  • Application of LCE3E to diagnosis and treatment of oral cancer
  • Application of LCE3E to diagnosis and treatment of oral cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Oral cancer and normal tissues were collected, grouped and numbered: 3 cases of normal tissue blocks (numbered N1, N2, N3) and 2 cases of cancer tissue blocks (C1, C2).

[0039] 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 Table 1 that all the samples in this study met the requirements of transcriptome sequencing.

[0040] Table 1 Spectrophotometric detection of oral c...

Embodiment 2

[0042] Example 2 High-throughput transcriptome sequencing and analysis

[0043] Sequencing platform: Illumina's HiSeq2500 high-throughput sequencing platform

[0044] Transcriptome sequencing and matching:

[0045] The two samples of cancerous tissue and normal tissue we analyzed came from 2 cases of oral cancer patients and 3 cases of healthy people. These two sets of samples were subjected to high-throughput cDNA sequencing. From these two tissues we get 4.85×10 7 ,4.46×10 7 read pairs. We used TopHat to match reads to the UCSC reference human genome (hg19), and the unique matched read pairs ranged from 4.04 × 10 7 to 4.49×10 7 The proportion of reads matching to Ensembl reference genes ranged from 81% to 89%. 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, it is about 1.13×10 8 ), and only 1% of the reads were mapped to rRNA, indicat...

Embodiment 3

[0051] Example 3 Cancer tissue and normal tissue LCE3E gene expression

[0052] 1. Materials and methods

[0053] 1. Materials

[0054] Oral cancer patients were hospitalized from 2010 to 2014. The cancer tissues of 49 patients with oral cancer and the normal tissues of 23 healthy people were taken as controls.

[0055] 2. Method

[0056] 2.1 Extraction of total RNA in oral cancer group and normal group

[0057] Extract the RNA of the oral cancer group and the normal group according to the instructions of Kangwei Century Ultrapure RNA Extraction Kit (UltrapureRNAKit (DNaseI), Cat. No. CW0597), prove the integrity of the RNA by gel electrophoresis, and measure the concentration and purity of the RNA with a nucleic acid protein analyzer .

[0058] Use the Ultrapure RNA Extraction Kit (Cat. No. CW0597) to extract, and the main steps are as follows:

[0059] 1. Sample processing, 30-50mg tissue is fully ground in liquid nitrogen and then added 1ml BufferRLT, or homogenized af...

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Abstract

The invention relates to application of LCE3E to diagnosis and treatment of an oral cancer. According to the application, transcriptome deep sequencing analysis is performed through a high-throughput sequencing platform to preliminarily screen out LCE3E genes obvious in differential expression in oral squamous cell carcinoma tissue and normal tissue, a further RT-PCR experiment proves high expression of the LCE3E genes in the oral squamous cell carcinoma tissue, and an siRNA interference experiment shows that interference to expression of the LCE3E genes can effectively suppress proliferation of oral cancer cells. Oral cancer molecular markers in the application have significant clinical practical application value.

Description

technical field [0001] The present invention relates to the technical field of bioengineering, in particular to the application of LCE3E in the diagnosis and treatment of oral cancer, and more specifically to the application of LCE3E in the diagnosis and treatment of oral squamous cell carcinoma. Background technique [0002] In today's society, cancer is one of the important lethal factors that threaten human health and life. According to the latest "World Cancer Report" statistics, in 2014, there were 14 million new cancer cases in the world, of which 3.07 million were in China, accounting for 21.8%. The number of cancer deaths in the world was about 8.2 million, of which 2.2 million were in China, accounting for 26.9%. When China accounts for the proportion of cancer in the world, the new cases and deaths of esophageal cancer account for 50% of the world, and it is the number one killer that endangers the health of Chinese people. The most common type, the prognosis is p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57484G01N33/68G01N2333/47
Inventor 宋宏涛靳晓霞杨承刚
Owner BEIJING MEDINTELL BIOMED CO LTD
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