New diagnosis and treatment target of nasopharynx cancer and application of new diagnosis and treatment target

A nasopharyngeal cancer, gene technology, applied in the field of molecular biology, can solve problems such as affecting the expression of target genes

Active Publication Date: 2016-02-24
李伟 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, miRNAs sometimes lead to histidine modification and DNA methylation in the promoter region, thereby affecting the expression of target genes.

Method used

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  • New diagnosis and treatment target of nasopharynx cancer and application of new diagnosis and treatment target
  • New diagnosis and treatment target of nasopharynx cancer and application of new diagnosis and treatment target
  • New diagnosis and treatment target of nasopharynx cancer and application of new diagnosis and treatment target

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] The collection of embodiment 1 sample and total RNA extraction

[0067] The tissues were all collected from the hospital from March 2011 to December 2013. The experimental group selected 7 patients with nasopharyngeal carcinoma confirmed by pathological examination, and the control group consisted of paracancerous tissues of the same sample.

[0068] experimental method:

[0069] 1 extraction method

[0070] 1) Take 80 mg tissue block, add 800 μl Lysis / Binding buffer, and use a homogenizer to homogenize the tissue block. During the homogenization process, the samples should be placed on ice to keep the cold state.

[0071] 2) Add 1 / 10 volume of HomogenateAdditive to the above homogenized tissue samples, and place on ice for 10 minutes.

[0072] 3) Add an equal volume of water-saturated phenol to the Lysis / Binding buffer, shake for 45 seconds, and centrifuge at 10,000×g for 5 minutes at room temperature.

[0073] 4) Carefully remove the supernatant into a new test tu...

Embodiment 2

[0083] Example 2 Sequencing and Data Analysis

[0084] The sequencing company carried out the establishment of the sequencing library and on-machine sequencing, and the sequencer used was the HiSeq2000 sequencer of Illumina Company.

[0085] According to the data analysis results provided by the company: 7 cases of nasopharyngeal carcinoma group and para-cancer group were statistically analyzed, the P value was less than 0.05, and the difference between the tumor group and the para-cancer control group was at least more than 2 times. Differentially expressed miRNAs were artificially selected and filtered to enter the VPS11 gene with significantly down-regulated expression and has-mir-632 with significantly up-regulated expression into our research scope, and the predicted target gene of has-mir-632 was predicted by software to be VPS11.

Embodiment 3

[0086] Example 3 Real-time PCR detection of the expression of VPS11 gene and miR-632 in nasopharyngeal carcinoma tissue

[0087] 1 sample collection:

[0088] 63 nasopharyngeal carcinoma tissues and adjacent tissues were obtained from the hospital (collection time from March 2011 to December 2013). The pathological properties and histological differentiation degree of 63 nasopharyngeal carcinoma were determined by two pathologists.

[0089] 2 RNA extraction

[0090] 2.1 miRNA extraction:

[0091] Removal of RNase for related experimental items:

[0092] ① Rinse and soak all glassware with DEPC before use, 120°C for 20 minutes under high pressure, and 180°C for more than 2 hours.

[0093] ②Plastic utensils (such as: EP tubes / tips) need to be soaked in 0.1% DEPC water overnight before use, and then the liquid is controlled to dry, 120°C and high pressure for 20 minutes, and dried in an oven for later use.

[0094] (1) Take out the frozen tumor tissue from liquid nitrogen, we...

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Abstract

The invention relates to a new diagnosis and treatment target of nasopharynx cancer and application of the new diagnosis and treatment target, and more particularly relates to a VPS11 gene and application that the VPS11 regulates and controls miRNA in treatment of nasopharynx cancer. Nasopharynx cancer tissues and para-carcinoma tissues are analyzed through high-throughput sequencing, mRNA and miRNA transcriptome information is obtained, after analysis, VPS11 gene and miR-363 are screened out and selected to perform fluorescent quantitation PCR verification and target verification, a result shows that the VPS11 gene and miR-363 are closely related to nasopharynx cancer, and VPS11 is a target gene of miR-632 in nasopharynx cancer. Through the invention, a new target is provided for clinical diagnosis and prevention detection of nasopharynx cancer, and has very good application value.

Description

technical field [0001] The present invention relates to the field of molecular biology, in particular to a new target for diagnosis and treatment of nasopharyngeal carcinoma and its application, and more specifically to the application of VPS11 gene and its regulating miRNA in the diagnosis and treatment of nasopharyngeal carcinoma. Background technique [0002] Nasopharyngeal carcinoma (Nasopharyngeal carcinoma, NPC) is one of the common malignant tumors of the head and neck in southern my country. The incidence rate of men is about twice that of women, and the highest incidence period is 40-50 years old. The incidence of nasopharyngeal carcinoma is related to genetic factors (genetic susceptibility), Epstein-Barr virus infection, environmental factors, eating habits and other factors. Early diagnosis and early treatment are the most effective means to save patients' lives and improve their quality of life. Unfortunately, nasopharyngeal carcinoma has an insidious onset and ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K31/713A61K45/06C12Q1/68G01N33/68A61P35/00
Inventor 李伟马会平
Owner 李伟
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