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Molecular markers for early detection of thyroid cancer and application of molecular markers

A technology for molecular markers and thyroid cancer, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve the problems of poor curative effect and no effect of thyroid cancer

Inactive Publication Date: 2019-04-19
思泰得精准(北京)医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both radiotherapy and chemotherapy have poor or no effect on thyroid cancer

Method used

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  • Molecular markers for early detection of thyroid cancer and application of molecular markers
  • Molecular markers for early detection of thyroid cancer and application of molecular markers
  • Molecular markers for early detection of thyroid cancer and application of molecular markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Screening for Gene Markers Related to Thyroid Cancer

[0043] 1. Sample collection

[0044] From October 2014 to January 2018, 25 tissue samples were obtained from the Department of Endocrinology, Beijing Luhe Hospital, all of which were confirmed by pathological examination, including 8 paracancerous tissue samples and 17 thyroid cancer samples, numbered Store in a -80°C low temperature refrigerator. All the above specimens were obtained with the consent of the organizational ethics committee.

[0045] 2. Total RNA extraction from tissue samples

[0046] use Reagent (invitrogen, Cat. No. 15596-018) was used to extract sample RNA, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0047] After collecting the samples, freeze them in liquid nitrogen. After taking them out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered:

[0048] ① Add Trizol and...

Embodiment 2

[0058] Example 2 Real-Time PCR Verification of FYB1 and CARMIL2 Expression in Thyroid Cancer Tissue and Paracancerous Tissue

[0059] 1. Materials

[0060] 21 cases of thyroid cancer tissue samples and 8 cases of paracancerous tissue samples were selected and grouped and numbered. All samples were confirmed by pathological examination.

[0061] 2. Method

[0062] 2.1 Extract the total RNA from the tissue sample, the same as the extraction method in Example 1.

[0063] 2.2 Synthesis of cDNA by reverse transcription

[0064] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0065] Using a reverse transcription kit, 1 μg of total RNA was reverse-transcribed with reverse transcription buffer to synthesize cDNA. A 25 μL reaction system was used, and 1 μg of total RNA was used as template RNA for each sa...

Embodiment 3

[0083] Example 3 kit assembly

[0084] Based on the primers obtained in Example 2, the kit for detecting thyroid cancer of the present invention was assembled, and the kit included primers for specifically amplifying the FYB1 and CARMIL2 genes, as shown in SEQID NO.1-SEQID NO.4.

[0085] The kit of the present invention also includes a pair of primers for specifically amplifying the housekeeping gene (GAPDH): shown in SEQ ID NO: 5 and SEQ ID NO: 6; also includes a SYBR Green polymerase chain reaction system, such as PCR buffer, SYBR Green fluorescent Dyes, dNTPs. The composition of described PCR buffer is 25mM KCl, 2.5mM MgCl 2 , 200mM (NH 4 ) 2 SO 4 . Through the optimization of primer concentration and annealing temperature, the reaction system was finally determined as follows:

[0086]

[0087] The optimal reaction conditions are: pre-denaturation at 95°C for 5 min, (denaturation at 95°C for 15 sec, annealing at 60°C for 45 sec, extension at 72°C for 35 sec) x 40 ...

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Abstract

The invention discloses molecular markers for early detection of thyroid cancer. FYB1 and CARMIL2 are adopted as the molecular markers, and the molecular markers are related to the occurrence of thyroid cancer. The invention also provides a thyroid cancer detection kit which can judge whether or not a subject suffers from thyroid cancer or has the risk of getting thyroid cancer by detecting the expression of FYB1 and CARMIL2 in the subject. Therefore, an instruction is provided for a clinician to offer a prophylactic or therapeutic scheme to the subject. In order to further study the etiologyand pathogenesis of thyroid cancer, a new direction is provided for exploring drug targets used for early prevention and treatment of thyroid cancer.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a molecular marker for early detection of thyroid cancer and its application. Background technique [0002] The thyroid gland is an important endocrine organ in the human body, secreting hormones related to metabolism such as heart rate, blood pressure, body temperature and weight. Thyroid cancer (thytoid carcinoma, TC) is a common malignant endocrine system tumor of the head and neck developed from thyroid follicular cells, accounting for about 3% of the total number of malignant tumors in the whole body, accounting for the first place in the incidence of malignant tumors of the head and neck , mainly occurs in young adults, is common in people aged 45-54, and is more common in women over the age of 45, with a male to female ratio of about 1:3. In recent years, the incidence of thyroid cancer has shown an upward trend. In 2012, more than 40,000 women and more than 10,000 men in the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/158
Inventor 纪智礼赵东熊慧姚广印李丽杨龙艳孙荣欣
Owner 思泰得精准(北京)医学检验实验室有限公司
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