Application of molecular marker stac2 in oral squamous cell carcinoma

A molecular marker, squamous cell carcinoma technology, applied in the field of biomedicine, can solve the problems of incompletely clear pathogenesis of oral squamous cell carcinoma and no molecular markers of oral squamous cell carcinoma

Active Publication Date: 2019-07-12
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the pathogenesis of oral squamous cell carcinoma is not completely clear, and there are no effective molecular markers for the diagnosis and treatment of oral squamous cell carcinoma in clinical practice. Finding new molecular markers is important for the study of the mechanism of oral squamous cell carcinoma and Clinical application is of great significance

Method used

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  • Application of molecular marker stac2 in oral squamous cell carcinoma
  • Application of molecular marker stac2 in oral squamous cell carcinoma
  • Application of molecular marker stac2 in oral squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Screening for Gene Markers Related to Oral Squamous Cell Carcinoma

[0061] 1. Sample collection

[0062] 6 cases of surrounding normal mucosa tissue and oral squamous cell carcinoma tissue were collected, all of which were confirmed by pathological diagnosis. All patients did not receive any form of treatment before operation. The surgically resected samples were frozen in liquid nitrogen, and the patients gave informed consent, and all the above-mentioned samples were obtained with the consent of the organizational ethics committee.

[0063] 2. Preparation of RNA samples (operated using QIAGEN tissue RNA extraction kit)

[0064] Take out the tissue samples frozen in liquid nitrogen, put the tissue samples into a pre-cooled mortar for grinding, and extract and isolate RNA according to the instructions in the kit. details as follows:

[0065] 1) Add Trizol and place at room temperature for 5 minutes;

[0066] 2) Add 0.2ml of chloroform, vibrate the centrif...

Embodiment 2

[0079] Example 2 QPCR sequencing to verify the differential expression of the STAC2 gene

[0080] 1. Large-sample QPCR verification of differential expression of STAC2 gene. According to the sample collection method in Example 1, 80 cases of normal mucosal tissues and 80 cases of oral squamous cell carcinoma tissues were selected.

[0081] 2. The RNA extraction steps are as described in Example 1.

[0082] 3. Reverse transcription:

[0083] 1) Reaction system:

[0084] Table 1 Reverse transcription reaction system

[0085]

[0086] 2) Reverse transcription reaction conditions

[0087] According to the reverse transcription reaction conditions in RNA PCR Kit (AMV) Ver.3.0.

[0088] 60min at 42°C, 2min at 99°C, 5min at 5°C.

[0089] 3) Polymerase chain reaction

[0090] 1) Primer design

[0091] QPCR amplification primers were designed according to the coding sequences of STAC2 gene and GAPDH gene in Genebank, and synthesized by Biomed Biotech. The specific primer se...

Embodiment 3

[0106] Example 3 Differential expression of STAC2 gene in oral squamous cell carcinoma cell lines

[0107] 1. Cell culture

[0108] Oral squamous cell carcinoma cell lines Tca8113 and HN13, and normal mucosal epithelial cell line HIOEC were purchased from the Ninth People's Hospital Affiliated to Shanghai Jiaotong University. The medium of HIOEC is K-SFM; the medium of Tca8113 and HN13 is DMEM; the culture medium containing 10% fetal bovine serum and 1% P / S was incubated at 37°C and 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0109] 2. Extraction of total cellular RNA

[0110] 1) Stop the culture when the cells reach 80-90% confluency, digest with 0.25% trypsin and collect the cells in 1.5ml EP tubes, add lm1Trizol to each tube and shake slowly to break the cells, place on ice for 10min.

[0111] 2) Remove protein and DNA: add 0.2ml ch...

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Abstract

The invention discloses application of a molecular marker STAC2 to oral squamous cell carcinoma. Experimental results show that STAC2 is highly expressed in a patient with oral squamous cell carcinoma; and a STAC2-specific siRNA for silencing STAC2 so as to reduce expression of STAC2, so proliferation of oral squamous cell carcinoma is inhibited. Thus, STAC2 can be applied to research on the pathogenesis of oral squamous cell carcinoma and is applicable as a molecular target to clinical diagnosis and treatment of oral squamous cell carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of molecular marker STAC2 in oral squamous cell carcinoma. Background technique [0002] Oral squamous cell carcinoma (OSCC) is a common malignant tumor in oral and maxillofacial surgery. Because of its high degree of malignancy and prone to lymph node metastasis, the prognosis is poor, and the deformity of the maxillofacial region is often secondary to surgery. It is very difficult for doctors to formulate treatment plans in clinical practice. have a great impact on the quality of life of patients. In my country, squamous cell carcinoma mostly occurs in adults aged 40-60, and more men than women, with tongue, cheek, gingiva, palate and maxillary sinus being the most common sites. Squamous cell carcinoma often metastasizes to regional lymph nodes, and distant metastasis may occur in advanced stages. [0003] Tumor is a disease at the molecular level, and tumor research ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/57484
Inventor 杜海威边洋孙耀兰
Owner BEIJING MEDINTELL BIOMED CO LTD
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