CMYA5 gene and expressed product of CMYA5 gene in application of laryngeal squamous cell carcinoma

A technology for expressing products, laryngeal squamous cell carcinoma, applied in the field of biomedicine, can solve the problem of unclear pathogenesis

Active Publication Date: 2017-11-17
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] With the in-depth research in the field of carcinogenesis and the continuous development of molecular biology, it has been found that many genes are abnormally expressed in laryngeal cancer tissues, but the specific pathogenesis is still unclear

Method used

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  • CMYA5 gene and expressed product of CMYA5 gene in application of laryngeal squamous cell carcinoma
  • CMYA5 gene and expressed product of CMYA5 gene in application of laryngeal squamous cell carcinoma
  • CMYA5 gene and expressed product of CMYA5 gene in application of laryngeal squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Screening for gene markers associated with laryngeal squamous cell carcinoma

[0070] 1. Sample collection

[0071] 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.

[0072] 2. Preparation of RNA samples

[0073] 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;

[0074] 2) Centrifuge at 12000rpm at 4°C for 5min, and transfer the supernatant to a 1.5ml RNase free EP tube;

[0075] 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;

[0076] 4) Centrifuge at 12000g for 15min at 4°C, centrifuge the solution into three lay...

Embodiment 2

[0092] Example 2 QPCR sequencing verification of differential expression of CMYA5 gene

[0093] 1. Large-sample QPCR verification of differential expression of CMYA5 gene. 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.

[0094] 2. The specific steps of RNA extraction are as described in Example 1.

[0095] 3. Reverse transcription

[0096] 3. Reverse transcription: FastQuant cDNA First Strand Synthesis Kit (Product No.: KR106) was used for mRNA reverse transcription. Specific steps are as follows:

[0097] (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;

[0098] (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...

Embodiment 3

[0113] Example 3 Differential expression of CMYA5 protein detected by protein immunoblotting

[0114] 1. Extraction of total tissue protein

[0115] 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.

[0116] 2. Determination of total protein concentration

[0117] The protein concentration was determined according to the instructions of the BCA protein concentration determination kit.

[0118] 3. SDS-PAGE electrophoresis

[0119] Prepare 8% separating gel and 5% stacking gel according to the instructions of the SDS-PAGE gel preparation kit and perform electrophoresis.

[0120] 4. Western detection

[0121] 1) Electrotransfer

[0122] Put the PVDF membrane into methanol ...

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Abstract

The invention discloses a CMYA5 gene and an expressed product of the CMYA5 gene in application of a laryngeal squamous cell carcinoma. Low expression of CMYA5 is presented in a laryngeal squamous cell carcinoma tissue. By increasing the expression of the CMYA5, cell proliferation and invasion of the laryngeal squamous cell carcinoma can be reduced. The sensibility and specificity of diagnosing of the laryngeal squamous cell carcinoma are greatly improved, and meanwhile a new target spot is provided for precise treatment of the laryngeal squamous cell carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of CMYA5 gene and its expression product in laryngeal squamous cell carcinoma, in particular to the application in the diagnosis and treatment of laryngeal squamous cell carcinoma. Background technique [0002] Laryngeal cancer is one of the most common head and neck tumors in the world, accounting for approximately 6% of all tumors. Worldwide, the average number of new cases of laryngeal cancer exceeds 500,000 per year, and the number of new deaths exceeds 200,000 per year. The 5-year survival rate is about 50%-70%. Although there have been a lot of advances in surgery, radiotherapy and chemotherapy in recent years, the long-term prognosis of patients with laryngeal cancer is still unsatisfactory. Tumor local recurrence, lymph node metastasis and distant metastasis are the main reasons affecting the prognosis of patients with laryngeal cancer. The malignant biological b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K48/00A61K38/17A61K45/00A61P35/00
CPCA61K38/1709A61K45/00A61K48/005C12Q1/6886C12Q2600/136C12Q2600/158G01N33/57407G01N33/57484G01N2333/47
Inventor 台德强常鹏
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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