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ANKRD1 as diagnosis and treatment target of tongue squamous cell carcinoma

A tongue squamous cell carcinoma, gene technology, applied in the treatment, ANKRD1 gene in the field of diagnosis of tongue squamous cell carcinoma, can solve the problem that the molecular mechanism is not clear

Inactive Publication Date: 2017-06-30
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although researchers have successively discovered that dozens of oncogenes or tumor suppressor genes are involved in the occurrence and development of tongue cancer, so far, the molecular mechanism of tongue cancer has not been clarified, and there may be other tumor-related genes involved. The pathogenesis of tongue cancer

Method used

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  • ANKRD1 as diagnosis and treatment target of tongue squamous cell carcinoma
  • ANKRD1 as diagnosis and treatment target of tongue squamous cell carcinoma
  • ANKRD1 as diagnosis and treatment target of tongue squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Differential expression of ANKRD1 gene

[0066] 1. Experimental materials

[0067] 5 tissue samples of tongue squamous cell carcinoma were obtained from oral and maxillofacial surgery patients, including 2 cases of well-differentiated squamous cell carcinoma, 2 cases of moderately differentiated squamous cell carcinoma, and 1 case of poorly differentiated squamous cell carcinoma; including 2 males and 3 females. At the same time, each case of cancerous tissue was selected as the normal tissue at >5cm around the cancerous tumor as its own control. All patients had not undergone chemotherapy, radiotherapy, biological therapy and other treatments for tumors before seeing the doctor. A part of the tissue was immediately stored in liquid nitrogen after collection.

[0068] 2. RNA extraction and cDNA synthesis

[0069] Total RNA was extracted with Trizol RNA reagent (Invitrogen Company), and total RNA was identified by UV spectrophotometer (ND-1000, NanoDrop Compan...

Embodiment 2

[0078] Example 2 Verification of differentially expressed genes in large samples

[0079] 1. Research object

[0080] According to the method of Example 1, 45 cases of tongue squamous cell carcinoma tissue samples and 50 cases of normal tissue samples were collected.

[0081] 2. RNA extraction and cDNA synthesis

[0082] RNA extraction and cDNA synthesis were performed according to the method in Example 1.

[0083] 3. RT-PCR

[0084] The primers were designed by the primer design software Primer 5.0, and synthesized by Dalian Bao Biological Company and Shanghai Yingjun Company. The primer sequences used for ANKRD1 gene and internal reference gene are as follows:

[0085] ANKRD1 gene primer sequence

[0086] 5'-AGACAGAGAAGGAGATAC-3' (SEQ ID NO.1),

[0087] 5'-GCCATACATAATCAGGAG-3' (SEQ ID NO.2);

[0088] GAPDH gene primer sequence

[0089] 5'-AAGGTCGGAGTCAACGGATTTG-3' (SEQ ID NO.3),

[0090] 5'-CCATGGGTGGAATCATATTGGAA-3' (SEQ ID NO. 4).

[0091] 1 μl of cDNA product w...

Embodiment 3

[0100] Example 3 Determination of the expression of ANKRD1 gene on the proliferation ability of tongue squamous cell carcinoma cells

[0101] 1. Interfering with ANKRD1 gene expression

[0102] 1.1 siRNA synthesis

[0103] Design and synthesize siRNA sequence siRNA-ANKRD1 according to the mRNA sequence of ANKRD1 gene:

[0104] The sense strand is 5'-ACAAACAUCUGGAUUGUUCUU-3' (SEQ ID NO.5);

[0105] The antisense strand is 5'-GAACAAUCCAGAUGUUUGUGA-3' (SEQ ID NO.6),

[0106] The above siRNA sequences and the negative control siRNA sequence (siRNA-NC) (the negative control siRNA has no homology to the sequence of the ANKRD1 gene) were provided by Shanghai Gemma Pharmaceutical Technology Co., Ltd.

[0107] 1.2 Culture and transfection of tongue squamous cell carcinoma cells

[0108] Human tongue squamous carcinoma cell line HN4 was cultured in DMEM / F12 medium containing 10% FBS, 100 U / mL penicillin and 100 μg / mL streptomycin. All cells were placed in 5% CO 2 in a 37 °C cell c...

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Abstract

The invention discloses ANKRD1 as a diagnosis and treatment target of tongue squamous cell carcinoma. Accordingly, ANKRD1 can be used for developing tongue squamous cell carcinoma diagnosis products and tongue squamous cell carcinoma treatment drugs. By means of research results, theoretical basis is provided for personalized treatment plan making by clinicians, and the novel drug target can be provided for development of the tongue squamous cell carcinoma treatment drugs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of ANKRD1 gene in the diagnosis and treatment of squamous cell carcinoma of the tongue. Background technique [0002] Tongue cancer is one of the most common malignant tumors in the oral and maxillofacial region. Squamous cell carcinoma is the most common and ranks first among oral cancers. Tongue cancer mostly occurs on the side edge of the middle third of the tongue. It has a high degree of malignancy and strong local infiltration. It often involves the tongue muscle, restricts tongue movement, and affects speech, eating, and swallowing functions. Despite great progress in treatment, the survival rate of tongue cancer patients has not improved significantly in the past few decades. [0003] Current studies have shown that smoking, drinking and other chemical factors, human papillomavirus (human papillomavirus, HPV) and genetic factors are closely related to the occ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
Inventor 肖枫杜海威
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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