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Diagnosis and treatment markers of hypopharyngeal carcinoma and its application

A pharyngeal cancer and material technology, applied in the field of tumor diagnosis, treatment, and prognosis prediction, can solve the problems of swallowing organ dysfunction, affecting the quality of life of patients, and many complications

Active Publication Date: 2019-09-10
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the particularity of the growth site of hypopharyngeal cancer, after surgical treatment, it may lead to dysfunction of language, breathing and swallowing organs, seriously affecting the quality of life of patients; while radiotherapy and chemotherapy have incomplete treatment, many complications, and more pain for patients. Major and other issues

Method used

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  • Diagnosis and treatment markers of hypopharyngeal carcinoma and its application
  • Diagnosis and treatment markers of hypopharyngeal carcinoma and its application
  • Diagnosis and treatment markers of hypopharyngeal carcinoma and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1 Gene Chip Screening for Differentially Expressed Genes

[0072] 1. Materials:

[0073] Eight patients with primary hypopharyngeal carcinoma underwent surgical resection of cervical lymph node dissection at the same time, and the normal hypopharyngeal mucosa tissues of nine patients with non-hypopharyngeal carcinoma were taken as controls. All cancer tissues were pathologically confirmed as hypopharyngeal carcinoma. All patients with primary hypopharyngeal carcinoma did not receive radiotherapy and chemotherapy before operation, and the clinical data of all cases were complete.

[0074] 2. Obtaining tissue RNA

[0075] Total tissue RNA was extracted using the Trizol one-step method.

[0076] 3. Determination of RNA purity and concentration

[0077] Take 1 μl of RNA solution, measure OD260 and OD280 with the instrument, the RNA concentration is OD260 value × dilution factor × 40 / 1000, calculate OD260 / OD280, the ratio of 1.7-2.0 means that the RNA solution ha...

Embodiment 2

[0091] Example 2 Large sample validation of differentially expressed genes screened out

[0092] Based on the results of the previous high-throughput transcriptome deep sequencing, and according to the size of the P value, we selected the SCLY gene for verification.

[0093] 1. Sample collection

[0094] According to the method of Example 1, 45 cases of hypopharyngeal cancer tissues and 50 cases of normal control tissues were collected.

[0095] 2. Validation at the mRNA level

[0096] 2.1 Extract tissue RNA

[0097] Step is with embodiment 1.

[0098] 2.2 Reverse transcription

[0099] Reverse transcription using Primescript 1 st strand cDNA synthesis kit kit, the operation steps are as follows:

[0100] (1) Add the following reaction liquid in the microcentrifuge tube, as shown in Table 1:

[0101] Table 1 Reaction liquid

[0102] Reagent dose RNA 2.0μg dNTP 1.0μl Oligo(dT) 2.0μl RNase free dH 2 o

Add to 10.0μl

[010...

Embodiment 3

[0128] Example 3 SCLY gene overexpression

[0129] 1. Plasmid construction

[0130] Amplification primers are designed according to the coding sequence of SCLY gene, and the design of primers is well known to those skilled in the art. Amplify the coding sequence of the full-length SCLY 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 -SCLY was used in subsequent experiments.

[0131] 2. Culture and transfection of hypopharyngeal carcinoma cells

[0132] 2.1 Cell culture

[0133] Hypopharyngeal cancer FADU cells were placed in RPMI 1640 medium containing 10% fetal bovine serum (FBS), penicillin 100 U / ml, and streptomycin 100 μg / ml at 37°C, 5% CO 2 , cultivated in a saturated humidity environment.

[0134] 2.2 Cell transfection

[0135] (1) The day before transfection, 0.5-2*10 5 Tumor cells wer...

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Abstract

The invention discloses a selenocysteine lyase (SCLY) gene, which can be taken as a molecular marker for hypopharyngeal carcinoma diagnosis. As found by a gene chip and a QPCR (Quantitative Polymerase Chain Reaction) experiment, a remarkable difference exists between expressions of the SCLY gene and an encoding protein of the SCLY gene in normal contrast tissues and hypopharyngeal carcinoma tissues, namely, the SCLY gene expression condition in the hypopharyngeal carcinoma tissues can be detected to judge whether a subject suffers from the hypopharyngeal carcinoma or has a risk of suffering from the hypopharyngeal carcinoma or not. According to the relevance, a kit for diagnosing the hypopharyngeal carcinoma is developed, and can be used for diagnosing the hypopharyngeal carcinoma by detecting the expression of the SCLY gene. The diagnosis kit provided by the invention can be applied to early diagnosis of diseases, and has a wide clinical application prospect. Moreover, the invention further discloses a SCLY gene which can be taken as a molecular target for treating the hypopharyngeal carcinoma, thereby laying a basis for the development work of medicaments for treating the hypopharyngeal carcinoma.

Description

technical field [0001] The present invention relates to the fields of tumor diagnosis, treatment and prognosis prediction, more specifically, the present invention relates to a method for tumor diagnosis and prognosis prediction by means of detecting SCLY abnormality; and a tumor therapeutic agent for activating SCLY gene or protein. Background technique [0002] Hypopharyngeal squamous cell carcinoma is one of the most common malignant tumors in otolaryngology head and neck surgery. It has the characteristics of hidden location, strong infiltration, easy submucosal spread, and multicentric growth of primary lesions. Due to the absence of obvious symptoms in the early stage, the lack of specific signs and reliable diagnostic measures, most patients are already in the late stage when they are clinically diagnosed and treated, and the lesions often involve organs such as the laryngeal cavity, cervical esophagus, and tongue base; at the same time, Due to the abundant lymphatic ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574A61K45/00A61P35/00
Inventor 杨承刚董东杜海威
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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