Application of ASPRV1 used as biomarker in diagnosis and treatment of LSCC (laryngeal squamous cell cancer)

A technology for laryngeal squamous cell carcinoma and drugs, which is applied in the determination/testing of microorganisms, biomaterial analysis, biochemical equipment and methods, etc., can solve problems such as unclear etiology, and achieve the effect of improving accuracy

Active Publication Date: 2017-09-15
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the pathogenesis of laryngeal cancer is considered to be related to bad living habits (smoking, drinking), adult la

Method used

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  • Application of ASPRV1 used as biomarker in diagnosis and treatment of LSCC (laryngeal squamous cell cancer)
  • Application of ASPRV1 used as biomarker in diagnosis and treatment of LSCC (laryngeal squamous cell cancer)
  • Application of ASPRV1 used as biomarker in diagnosis and treatment of LSCC (laryngeal squamous cell cancer)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Screening for gene markers associated with laryngeal squamous cell carcinoma

[0075] 1. Sample collection

[0076] Six cases of laryngeal squamous cell carcinoma tissues and corresponding normal mucosal tissue samples were collected, and the informed consent of the patients was obtained for the tissue samples, and the approval of the organizational ethics committee was obtained.

[0077] 2. Preparation of RNA samples

[0078] 1) Add liquid nitrogen to grind the tissue to powder, add 1ml TRIzol (Invitrogen) solution, mix by pipetting to fully lyse the tissue, and let stand for 5 minutes;

[0079] 2) Centrifuge at 12000rpm at 4°C for 5min, and transfer the supernatant to a 1.5ml RNase free EP tube;

[0080] 3) Add 200 μl of chloroform, vigorously shake and mix for 30 s, make the aqueous phase and organic phase fully contact, and let stand at room temperature for 15 min;

[0081] 4) Centrifuge at 12000g for 15min at 4°C, centrifuge the solution into three lay...

Embodiment 2

[0097] Example 2 QPCR sequencing to verify the differential expression of ASPRV1 gene

[0098] 1. Large-sample QPCR verification of ASPRV1 gene differential expression. According to the sample collection method in Example 1, 50 cases of normal paracancerous mucosa tissues and 50 cases of laryngeal squamous cell carcinoma tissues were selected from patients with laryngeal squamous cell carcinoma.

[0099] 2. The specific steps of RNA extraction are as described in Example 1.

[0100] 3. Reverse transcription

[0101] 3. Reverse transcription: FastQuant cDNA First Strand Synthesis Kit (Product No.: KR106) was used for mRNA reverse transcription. Specific steps are as follows:

[0102] (1) Add 5×gDNA Buffer 2.0μl, total RNA 1μg, add RNase Free ddH 2 O to bring the total volume to 10 μl, heat in a water bath at 42°C for 3 minutes;

[0103] (2) Construct a 20μl reaction system, 10× Fast RT Buffer 2.0μL, RT Enzyme Mix 1.0μl, FQ-RT Primer Mix 2.0μl, RNase Free ddH 2 O 5.0 μl wa...

Embodiment 3

[0120] Example 3 Differential expression of ASPRV1 protein detected by protein immunoblotting

[0121] 1. Extraction of total tissue protein

[0122] Cut the tissue with scissors and put it into a glass homogenizer placed in ice. Mix the RIPA lysate and PMSF at a ratio of 100:1, and add the corresponding amount of RIPA lysate, crush the tissue with a glass homogenizer until it is fully lysed, suck the lysed liquid into an EP tube, centrifuge at 14,000 rpm for 5 min at 4°C, and collect the supernatant.

[0123] 2. Determination of total protein concentration

[0124] The protein concentration was determined according to the instructions of the BCA protein concentration determination kit.

[0125] 3. SDS-PAGE electrophoresis

[0126] Prepare 8% separating gel and 5% stacking gel according to the instructions of the SDS-PAGE gel preparation kit and perform electrophoresis.

[0127] 4. Western detection

[0128] 1) Electrotransfer

[0129] Put the PVDF membrane into methanol...

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Abstract

The invention discloses an application of ASPRV1 used as a biomarker in diagnosis and treatment of LSCC (laryngeal squamous cell cancer). Expression up-regulation of ASPRV1 in LSCC tissue is discovered with a high-throughput sequencing technology, significant overexpression of the gene and protein expressed by the gene is further verified through experiments, and the ASPRV1 can be used as the diagnosis marker to be applied to clinical diagnosis of the LSCC. In-vitro cell experiments further prove that change of the expression level of the ASPRV1 can play a role in inhibiting proliferation, migration and invasion of cells, and the ASPRV1 can be used as a potential target to be applied to clinical treatment of the LSCC.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of ASPRV1 as a biomarker in the diagnosis and treatment of laryngeal squamous cell carcinoma. Background technique [0002] Laryngeal cancer is one of the most common malignant tumors in otolaryngology head and neck surgery, accounting for 5.7%-7.6% of systemic cancers. 96% to 98% of laryngeal cancers are squamous cell carcinomas, and other pathological types of carcinomas such as adenocarcinoma and sarcoma are less common. Laryngeal cancer is more likely to occur in the forty to sixty years old, and the majority of male patients. In recent years, with the continuous development of economy, the deterioration of living environment and the change of people's lifestyle, the incidence of laryngeal cancer is gradually increasing. Surgical resection of the tumor is the preferred treatment for laryngeal cancer. With the cooperation of radiotherapy, chemotherapy, and gene-target...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/574G01N33/573A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/136C12Q2600/158G01N33/573G01N33/57407G01N33/57484G01N2333/96472
Inventor 常鹏台德强
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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