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Diagnostic marker and therapeutic target for laryngeal squamous cell carcinoma

A technology of laryngeal squamous cell carcinoma and drugs, applied in the field of diagnostic markers and therapeutic targets of laryngeal squamous cell carcinoma, can solve the problems of unclear pathogenesis, intractable treatment, recurrence or metastasis, etc.

Inactive Publication Date: 2017-11-03
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 development of biotechnology, people have a certain understanding of the occurrence and development of laryngeal squamous cell carcinoma, but its specific pathogenesis is still unclear
Although laryngeal cancer can be treated comprehensively by various methods such as surgery, chemotherapy, and radiotherapy, there are still some patients who, despite radical surgery, radiotherapy, and chemotherapy, still die due to local recurrence or metastasis.

Method used

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  • Diagnostic marker and therapeutic target for laryngeal squamous cell carcinoma
  • Diagnostic marker and therapeutic target for laryngeal squamous cell carcinoma
  • Diagnostic marker and therapeutic target for laryngeal squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Screening for gene markers associated with laryngeal squamous cell carcinoma

[0066] 1. Sample collection

[0067] 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.

[0068] 2. Preparation of RNA samples

[0069] 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;

[0070] 2) Centrifuge at 12000rpm at 4°C for 5min, and transfer the supernatant to a 1.5ml RNase free EP tube;

[0071] 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;

[0072] 4) Centrifuge at 12000g for 15min at 4°C, centrifuge the solution into three lay...

Embodiment 2

[0088] Example 2 QPCR sequencing to verify the differential expression of MYOZ3 gene

[0089] 1. Large-sample QPCR verification of differential expression of MYOZ3 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.

[0090] 2. The specific steps of RNA extraction are as described in Example 1.

[0091] 3. Reverse transcription

[0092] 3. Reverse transcription: FastQuant cDNA First Strand Synthesis Kit (Product No.: KR106) was used for mRNA reverse transcription. Specific steps are as follows:

[0093] (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;

[0094] (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 w...

Embodiment 3

[0109] Example 3 Western blot assay to detect the differential expression of MYOZ3 protein

[0110] 1. Extraction of total tissue protein

[0111] 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.

[0112] 2. Determination of total protein concentration

[0113] The protein concentration was determined according to the instructions of the BCA protein concentration determination kit.

[0114] 3. SDS-PAGE electrophoresis

[0115] Prepare 8% separating gel and 5% stacking gel according to the instructions of the SDS-PAGE gel preparation kit and perform electrophoresis.

[0116] 4. Western detection

[0117] 1) Electrotransfer

[0118] Put the PVDF membrane into methanol so...

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Abstract

The invention discloses a diagnostic marker and a therapeutic target for laryngeal squamous cell carcinoma. The diagnostic marker and the therapeutic target for laryngeal squamous cell carcinoma are MYOZ3, and belong to the technical field of new application of genes. According to the diagnostic marker and the therapeutic target disclosed by the invention, down-regulation of expression of the marker MYOZ3 in patients suffering from the laryngeal squamous cell carcinoma is firstly found; moreover, through up-regulated expression of the MYOZ3, the proliferation of cancer cells can be reduced, and the MYOZ3 is prompted to be as a target, the MYOZ3 can be used for realizing accurate treatment on the laryngeal squamous cell carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a diagnostic marker and a therapeutic target of laryngeal squamous cell carcinoma. Specifically, the diagnostic marker and the therapeutic target are MYOZ3. Background technique [0002] Laryngeal cancer is divided into primary and secondary two. Primary laryngeal cancer refers to the tumor whose primary site is in the larynx, and squamous cell carcinoma (Laryngeal Squamous Cell Carcinoma, LSCC) is the most common, accounting for more than 90%. The occurrence of laryngeal cancer is gradually developed through multiple stages, from normal mucosa, hyperplasia and hypertrophy, mild dysplasia, moderate dysplasia, severe dysplasia, carcinoma in situ, invasive carcinoma to metastasis. [0003] With the development of biotechnology, people have a certain understanding of the occurrence and development of laryngeal squamous cell carcinoma, but its specific pathogenesis is still unclear. Althou...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886G01N33/57407G01N33/57484G01N33/68G01N2333/4703
Inventor 台德强常鹏
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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