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Gene relevant to papillary thyroid tumors

A papillary, genetic technology, applied in the field of molecular biology, can solve problems such as inability to recognize tumors

Active Publication Date: 2015-08-19
BOZHOU CITY THE NEW HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Due to the complexity of tumor genetics, traditional screening methods for somatic mutations cannot provide an overall understanding of tumors

Method used

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  • Gene relevant to papillary thyroid tumors
  • Gene relevant to papillary thyroid tumors
  • Gene relevant to papillary thyroid tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Mutation Sequencing Method

[0049] 1.1 Collection of tumor tissue samples from subjects

[0050] 1.2 Genomic DNA extraction

[0051] Prepare the mortar after autoclaving, dry it and pour it into liquid nitrogen to pre-cool; take an appropriate amount of tissue and grind it in the mortar, supplement liquid nitrogen during the period, grind it into powder and thaw it; collect the tissue in the mortar with 800 μl PBS solution Put it into a centrifuge tube (1.5ml); centrifuge at 12000rpm for 1min, take out and discard the supernatant. Then add 200 μl buffer GA and shake until thoroughly suspended.

[0052] Add 20 μl of proteinase K (20 mg / ml), mix well, incubate at 56°C for 2 hours, and shake once every 20 minutes until the tissue dissolves.

[0053] Add 200 μl GB buffer and mix by inverting, and keep warm at 70°C for 10 minutes until the solution becomes clear.

[0054] Add 200 μl of absolute ethanol and shake fully for 15 sec. At this time, a flocculent pr...

Embodiment 2

[0067] Example 2: GAS8-AS1 and other genes can be used as targets for screening or detection of papillary thyroid cancer.

[0068] 2.1 Experimental method

[0069] The present invention uses the genome-wide exome technology as described in Example 1 to perform targeted sequencing on 91 paired tissues (thyroid cancer tissues and peripheral blood samples) of papillary thyroid cancer patients to obtain gene mutations. Finally, GAS8-AS1 and other genes were identified as susceptibility genes for papillary thyroid cancer in Chinese Han population. The paired tissues of papillary thyroid cancer patients refer to the thyroid cancer tissues and peripheral blood samples of the patients. The 91 pairs of tissue samples of thyroid cancer patients were collected by the Cancer Hospital of Chinese Academy of Medical Sciences and Zhejiang Cancer Hospital.

[0070] 2.2 Experimental results

[0071] 2.21 Gene mutation frequency statistics

[0072] Next-generation sequencing was performed on ...

Embodiment 3

[0083] Example 3: GAS8-AS1 gene detection can be used as a target for screening or detection of papillary thyroid cancer.

[0084] GAS8-AS1 gene (NCBI-GeneID: 750), also known as C16orf3 gene (open reading frame 3 on chromosome 16), is located in intron 2 of GAS8 gene on human chromosome 16. The GAS8-AS1 gene does not contain any intronic sequences, and is transcribed in the opposite direction to the GAS8 gene to produce a long non-coding RNA (lncRNA), which cannot be translated into protein. At present, the biological functions of this gene and its transcripts are still unclear.

[0085] The gene sequence of GAS8-AS1 (NCBI Reference Sequence: NR_122031.1) is as follows:

[0086] 1acctgcagtc ccagctactg ggcagcctga agcagcagga tggtgtgaac ccaggaggtg

[0087] 61gagcttgcag tgagccgagg tcgcgccacc gcactccagc ctgggccaca cagcgagatt

[0088] 121ccgtcagaat cagttacttt tcgggcacag ccccaggcca cttactgtga gcctttttct

[0089] 181ttctcaacac cacattcccc acagggaaaa cacatttctc acctcaaaag aagacaagac ...

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Abstract

The invention relates to a gene relevant to papillary thyroid tumors and an application thereof. According to the base sequence of the gene, real-time and quantitative PCR (Polymerase Chain Reaction) primers are designed and synthesized; the expression level of long-chain non-coding RNA (Ribonucleic Acid) transcribed by the gene is detected in a papillary thyroid carcinoma clinical case specimen; the result shows remarkable reducing of the expression level of the long-chain non-coding RNA in papillary thyroid tumor tissues and the long-chain non-coding RNA of the gene silencing can remarkably promote the growth of thyroid cancer cells. The gene relevant to the papillary thyroid tumors is expected to prepare preparations used in papillary thyroid carcinoma auxiliary diagnosis, gene therapy, curative effect prediction or prognosis.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a gene related to papillary thyroid tumor and a PCR detection method thereof. Background technique [0002] Thyroid cancer is the most common endocrine tumor, and its incidence has increased rapidly in recent years. It has become one of the most common malignant tumors and the fastest growing solid tumor in recent years. Among them, 90% are papillary thyroid cancer (papillary thyroid cancer, PTC). Although the mortality rate of papillary thyroid cancer is relatively low compared with other malignant tumors, the metastasis rate of papillary thyroid cancer is very high, and the lymph node metastasis rate is as high as 30%-50%. If it can be diagnosed in time, the best timing of operation will be lost as a result, and the mortality rate will increase significantly, which is an important indicator of poor prognosis. [0003] Epidemiological evidence shows that in many area...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/68C12N15/11
Inventor 熊良钟
Owner BOZHOU CITY THE NEW HEALTH TECH CO LTD
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