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Diagnosis and treatment markers of tongue squamous cell carcinoma

A technology of tongue squamous cell carcinoma and products, applied in the treatment, PIGR gene in the field of diagnosis of tongue squamous cell carcinoma, can solve the problems such as unclear molecular mechanism

Active Publication Date: 2020-03-31
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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  • Diagnosis and treatment markers of tongue squamous cell carcinoma
  • Diagnosis and treatment markers of tongue squamous cell carcinoma
  • Diagnosis and treatment markers of tongue squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Differential expression of embodiment 1 PIGR gene

[0063] 1. Experimental materials

[0064] 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.

[0065] 2. RNA extraction and cDNA synthesis

[0066] Total RNA was extracted with Trizol RNA reagent (Invitrogen Company), and total RNA was identified by UV spectrophotometer (ND-1000, NanoDrop Com...

Embodiment 2

[0075] Example 2 Verification of Differentially Expressed Genes in Large Samples

[0076] 1. Research object

[0077] 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.

[0078] 2. RNA extraction and cDNA synthesis

[0079] RNA extraction and cDNA synthesis were performed according to the method in Example 1.

[0080] 3. RT-PCR

[0081] 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 PIGR gene and internal reference gene are as follows:

[0082] PIGR gene primer sequence

[0083] 5'-AACTATACAGGAAGAATA-3' (SEQ ID NO.3),

[0084] 5'-TATTACTATTGGAATCATC-3' (SEQ ID NO.4);

[0085] GAPDH gene primer sequence

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

[0087] 5'-CCATGGGTGGAATCATATTGGAA-3' (SEQ ID NO. 6).

[0088] 1 μl of cDNA product was ...

Embodiment 3

[0097] The expression of embodiment 3 PIGR gene is to the determination of the proliferative ability of tongue squamous cell carcinoma cell

[0098] 1. Interfering with PIGR gene expression

[0099] 1.1 siRNA synthesis

[0100] Design and synthesize siRNA sequence siRNA-PIGR according to the mRNA sequence of PIGR gene:

[0101] The sense strand is 5'-AGUACUUCAACAGAUCAUCAA-3' (SEQ ID NO.7);

[0102] The antisense strand is 5'-GAUGAUCUGUUGAAGUACUUA-3' (SEQ ID NO.8),

[0103] The above siRNA sequences and the negative control siRNA sequence (siRNA-NC) (the negative control siRNA has no homology to the sequence of the PIGR gene) were provided by Shanghai Gemma Pharmaceutical Technology Co., Ltd.

[0104] 1.2 Culture and transfection of tongue squamous cell carcinoma cells

[0105] Human tongue squamous carcinoma cell line HN4 was cultured in DMEM / F12 medium containing 10% FBS, 100 U / m L penicillin and 100 μg / m L streptomycin. All cells were placed in 5% CO 2 in a 37 °C cell ...

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Abstract

The invention discloses a PIGR (Polymeric Immunoglobulin Receptor) gene as a molecular marker for early diagnosis of tongue squamous carcinoma. Tests show that the PIGR gene expression in tongue squamous carcinoma tissue is remarkably increased when being compared with that in normal tissue, and RNA (Ribonucleic Acid) interference tests show that the PIGR is capable of influencing capability of proliferation, migration, invasion and tumor formation of tongue squamous carcinoma cells. According to research achievements of the invention, medicines for inhibiting PIGR gene expression can be developed, so that clinical prevention and treatment on tongue squamous carcinoma can be achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of PIGR 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 occurrence an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K48/00A61K31/713A61K39/395A61P35/00
CPCA61K31/713A61K39/39558C12Q1/6886C12Q2600/158G01N33/57407G01N33/68G01N2333/70535
Inventor 杨承刚杜海威
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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