Pancreatic neuroendocrine tumor susceptibility gene locus and detection method and kit

A kit and gene technology, applied in the fields of molecular biology and medicine, can solve the problems of unconfirmed reports on the correlation of YY1 gene with pancreatic neuroendocrine tumors, and unproven reports on the correlation of pancreatic neuroendocrine tumors

Active Publication Date: 2016-06-01
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although there have been some studies on the correlation between gene mutations and polymorphic sites and pancreatic neuroendocrine tumors, there is no report on the correlation between YY1 gene and functional pancreatic neuroendocrine tumors, let alone the YY1 gene mutation described in the present invention A report on the correlation between loci and pancreatic neuroendocrine tumors

Method used

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  • Pancreatic neuroendocrine tumor susceptibility gene locus and detection method and kit
  • Pancreatic neuroendocrine tumor susceptibility gene locus and detection method and kit
  • Pancreatic neuroendocrine tumor susceptibility gene locus and detection method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] 1.1 Clinical specimens

[0100] Diagnostic criteria for functional pancreatic neuroendocrine tumors: hypoglycemia and other related clinical symptoms, assessment of blood insulin and blood glucose levels (prolonged oral glucose tolerance test), CT / PET-CT impact on diagnosis, and pathological diagnosis of surgically resected tumors. Genomic DNA was extracted from islet cell tumors preserved in liquid nitrogen and matched peripheral blood samples, and from formalin-fixed paraffin-embedded (FFPE) insulinoma tissue samples. DNA was prepared using QIAGEN's DNeasy kit and QIA&DNAFFPE tissue kit.

[0101] Deparaffinize FFPE tissue using standard techniques. Immunofluorescence staining was performed on the samples according to standard protocol. The following primary antibodies were used: polyclonal rabbit anti-YY1 antibody (1:100; OriGene, clone EPR4651) and polyclonal guinea pig anti-insulin (1:400, Dako Corporation). Secondary antibodies for immunofluorescence staining we...

Embodiment 2

[0138] Pancreatic Neuroendocrine Tumor Gene Mutation Detection Kit

[0139] As described in Example 1, there is a nucleotide sequence of the YY1 gene: that is, the 1115th C→G mutation in SEQ ID NO.: 1 is closely related to pancreatic neuroendocrine tumor diseases. Therefore, specific primers for the YY1 gene can be designed based on this mutation and can be amplified using the patient's DNA as a template for detection.

[0140] Prepare a test kit (100 person-times), which contains:

[0141]

[0142] A test group composed of 100 individuals was randomly selected, including subjects who were unknown whether they had pancreatic neuroendocrine tumors, patients who were known to have pancreatic neuroendocrine tumors, and normal people who were tested to have no pancreatic neuroendocrine tumors.

[0143]Obtain a small amount of islet tissue samples from the subjects to be tested in the test group, and extract DNA using conventional methods. Dilute the PCR primers in the pancrea...

Embodiment 3

[0149] Adjunct detection of pancreatic neuroendocrine tumors

[0150] The test in Example 2 was repeated, with the difference that 80 people (whose symptoms of islet cell tumor were not known before the test) were randomly selected for the test.

[0151] Prepare a test kit (100 person-times), which contains:

[0152]

[0153] Take the islet cell tissue sample of the subject to be tested, and use conventional methods (or use a specific kit) to extract DNA from it. Dilute the PCR primers in the detection kit to 1 μmol / μl, and use the extracted DNA as a template to perform PCR reaction with the provided primers. After the PCR products were purified, they were sequenced with an ABI3730 DNA sequencer, and Polyphred software was used for sequence interpretation and SNV confirmation.

[0154] The results also confirmed that the proportion of islet cell tumors in the test subject containing the C→G mutation at position 100, 743, and 807 of chromosome 14 (C site at position 1115 i...

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Abstract

Disclosed in the invention are a pancreatic neuroendocrine tumour susceptibility gene loci and detection methods and kits. In particular, disclosed in the invention is a pancreatic neuroendocrine tumour susceptibility gene loci based on whole genome sequencing findings, the loci is a high frequency T372R somatic mutation of a YY1 (Yin Yang1) gene. Also disclosed in the invention are methods for the detection of pancreatic neuroendocrine tumour susceptibility and pancreatic neuroendocrine tumour subtype classification, and corresponding detection kits.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, it relates to the correlation between YY1 gene and its mutation sites and pancreatic neuroendocrine tumors, which can be used for typing diagnosis and pathogenic gene sites as drug targets in pancreatic neuroendocrine tumors. The present invention also relates to methods and kits for detecting these mutation sites. Background technique [0002] The major type of functional pancreatic neuroendocrine tumor is islet cell tumor. Islet cell tumors do not rely on glucose to stimulate continuous excessive secretion of insulin, which can cause clinical symptoms such as severe hypoglycemia. [0003] Although some genetic studies of pancreatic neuroendocrine tumor-associated genes have been carried out, and some researches have found some risk loci related to pancreatic neuroendocrine tumors in the past few years, the genetic background leading to sporadic functio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/156
Inventor 宁光曹亚南王卫庆王俊高志博李林
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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