WO2014190927A1 - Loci de gène de sensibilité tumorale pancréatique neuroendocrinienne et procédés et kits de détection - Google Patents

Loci de gène de sensibilité tumorale pancréatique neuroendocrinienne et procédés et kits de détection Download PDF

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WO2014190927A1
WO2014190927A1 PCT/CN2014/078833 CN2014078833W WO2014190927A1 WO 2014190927 A1 WO2014190927 A1 WO 2014190927A1 CN 2014078833 W CN2014078833 W CN 2014078833W WO 2014190927 A1 WO2014190927 A1 WO 2014190927A1
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gene
mutation
seq
pancreatic neuroendocrine
kit
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PCT/CN2014/078833
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English (en)
Chinese (zh)
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宁光
曹亚南
王卫庆
王俊
高志博
李林
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中国科学院上海生命科学研究院
上海交通大学医学院附属瑞金医院
深圳华大基因研究院
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Publication of WO2014190927A1 publication Critical patent/WO2014190927A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/112—Disease subtyping, staging or classification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the present invention relates to the fields of molecular biology and medicine. More specifically, it relates to the association of the YY1 gene and its mutation site with pancreatic neuroendocrine tumors, which can be used for typing diagnosis and pathogenic gene loci as drug targets in pancreatic neuroendocrine tumors. The invention also relates to methods and kits for detecting these mutation sites. Background technique
  • pancreatic neuroendocrine tumors The main type of functional pancreatic neuroendocrine tumors is islet cell tumor. Islet cell tumors do not rely on glucose stimuli to continue to secrete excessive amounts of insulin, which can cause clinical symptoms such as severe hypoglycemia.
  • pancreatic neuroendocrine tumor-related genes Although some genetic studies have been carried out on pancreatic neuroendocrine tumor-related genes, some research findings on pancreatic neuroendocrine tumors have been conducted in the past few years, but the genetic background for the dissemination of functional pancreatic neuroendocrine tumors is still Lack of understanding.
  • pancreatic neuroendocrine tumor susceptibility genes in order to diagnose, classify and treat pancreatic neuroendocrine tumors as early as possible, there is an urgent need in the art to find pancreatic neuroendocrine tumor susceptibility genes, and to develop methods, kits, or related methods for detecting pancreatic neuroendocrine tumor susceptibility genes. Treatment drugs and programs. Summary of the invention
  • Another object of the present invention is to provide a novel method of treating pancreatic neuroendocrine tumors.
  • a method for non-diagnostic detection of a sample of a somatic cell site mutation of the YY1 gene in vitro comprising the steps of -
  • nucleotide sequence of the YY1 gene corresponds to position 100, 743, 807 of human chromosome 14 (NCBI36/hgl8).
  • the gene-specific primer has the sequences shown in SEQ ID NO: 3 and 4.
  • the amplification product is 80-2000 bp in length and contains the 1115th position in SEQ ID NO: 1.
  • the detecting is the detection of pancreatic neuroendocrine tumor cells.
  • a kit for detecting a pancreatic neuroendocrine tumor comprising a primer for specifically amplifying a YY1 gene or a transcript, the primer having a length of 80-2000 bp and comprising the SEQ ID NO: amplification product at position 1 115 of 1.
  • the kit further comprises a reagent selected from the group consisting of:
  • the mutation is a single nucleotide mutation (SNV) :
  • the nucleotide sequence of the YY1 gene that is, the 1115th position in SEQ ID NO.: 1 C ⁇ G.
  • the primer has the sequence shown in SEQ ID NO: 3 and 4.
  • a YYi gene for the preparation of a reagent or kit for the diagnostic typing of pancreatic neuroendocrine tumors.
  • the reagent or kit is for detecting the following single nucleotide mutation (SNV) : the nucleotide sequence of the YY1 gene is the 1115th C ⁇ G in SEQ ID NO.: 1.
  • the reagent comprises a primer that specifically amplifies a YY1 gene or a transcript, an amplification product containing the mutation site, a probe that specifically binds to the mutation site, and specificity A nucleic acid chip that detects the mutation site.
  • the kit includes instructions for use and one or more of the following reagents - a container (a) and a primer for specifically amplifying the YY1 gene or transcript located in the container; And a probe located in the container that specifically binds to the mutation site; a container (c) and a nucleic acid chip located within the container that specifically detects the mutation site.
  • the nucleic acid chip further comprises for detecting additional pancreatic neuroendocrine The detection point of the tumor susceptible site.
  • the additional islet cell tumor somatic mutation site is selected from the group consisting of:
  • Missense mutation of the MLL3 gene chromosome 7, 151, 874, 40 (H3 ⁇ 4 A ⁇ G ;
  • Missense mutation of H3F3A gene chromosome 1, 226, 252, 135 ⁇ 6 ;
  • Frameshift mutation of LM02 gene chromosome 1, chromosome 33, 886, 330 insertion ((*/+0)
  • a polynucleotide molecule the molecule A primer comprising a specific amplification product of an amplification product containing a mutation site and/or a probe specifically binding to the SNV site, wherein the nucleotide molecule is used for preparing a neuroendocrine tumor of the pancreas
  • a kit for performing diagnostic typing and the mutation site is selected from the group consisting of the nucleotide sequence of the YY1 gene, that is, the 1115th C ⁇ G in SEQ ID NO.: 1.
  • the mutation site is the nucleotide sequence of the YY1 gene
  • the kit is for determining whether a patient with pancreatic neuroendocrine tumor is suitable for treatment with everolimus.
  • the pancreatic neuroendocrine tumor patient has the mutation site
  • the patient is indicated to be suitable for treatment with Everol imus.
  • the kit is for detecting pancreatic neuroendocrine tumor typing in a population of East Asia, particularly in the Chinese population.
  • a method of diagnostic typing of an individual pancreatic neuroendocrine tumor characterized in that it comprises the steps of:
  • the difference indicates that the individual has pancreatic neuroendocrine tumors belonging to the YY1 mutant subtype, with specific clinical features, prognosis and treatment.
  • the YY1 gene or transcript is detected and compared to the normal cellular DNA nucleotide sequence.
  • the YY1 mutant gene causes an up-regulation of the expression level of the downstream gene.
  • the downstream gene comprises IDH3A, UCP2, C0L1AK C0X5A, PGC-la, or PGC- ⁇ .
  • the downstream gene comprises IDH3A, UCP2, or C0L1A1.
  • the difference is the following mutation site:
  • the nucleotide sequence of the YY1 gene is the 1115th C ⁇ G in SEQ ID NO.: 1.
  • a YY1 gene for the preparation of a reagent or kit for determining whether a patient with a pancreatic neuroendocrine tumor is suitable for treatment with everolimus.
  • a polynucleotide molecule comprising a primer for an amplification product of a specific mutation site and/or a probe that specifically binds to the mutation site, Place
  • the nucleotide molecule is used for preparing a kit for determining whether a patient with pancreatic neuroendocrine tumor is suitable for treatment with everolimus, and the mutation site is selected from the group consisting of -
  • the nucleotide sequence of the YY1 gene is the 1115th C ⁇ G in SEQ ID NO.: 1.
  • pancreatic neuroendocrine tumor patient when the pancreatic neuroendocrine tumor patient has the mutation site, the patient is indicated to be suitable for treatment with Everol imus. It is to be understood that within the scope of the present invention, the above-described various technical features of the present invention and the technical features specifically described in the following (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here. DRAWINGS
  • Figure 1 shows the T372R hotspot mutation in the YY1 gene in functional pancreatic neuroendocrine tumors.
  • T372R mutation position based on the protein crystal structure of the YY1 protein zinc finger structure domain.
  • the T372R mutation is located in the functional domain of the third zinc finger structure and is on the surface bound to DNA, suggesting that it is involved in transcriptional regulation and DNA modification.
  • FIG. 2 shows the results of detection of the T372R hotspot mutation of the YY1 gene in islet cell tumors.
  • the T372R hotspot mutation of the YY1 gene in islet cell tumor cells was detected and determined by Sanger sequencing (a) and pyrosequencing (b) techniques.
  • Figure 3 shows the results of quantitative detection of YY1 gene expression in sporadic islet cell tumors. Mutant and wild type YY1 genes are expressed at higher levels in islet cell tumor cells.
  • Figure 4 shows the effect of the T372R mutation on the expression of downstream genes in the YY1 gene in islet cell tumors.
  • Figure 5 shows the strategy for constructing a transgenic mouse model.
  • the cell proliferation assay of Figure 6 showed that wild-type and mutant YY1 gene overexpression could induce significant proliferation of islet cell tumor cells, and T372R mutant YY1 gene promoted the proliferation of islet cell tumor cells.
  • Figure 7 shows that the expression of the islet cell proliferation marker Ki 67 was significantly increased in the YY1 transgenic mouse, and the increase in Ki67 after administration of the mTOR inhibitor was significantly inhibited.
  • the inventors have conducted extensive and intensive research to measure and analyze a large number of candidate genes. It was first discovered and demonstrated that the YY1 genome sequence is closely related to the occurrence of pancreatic neuroendocrine tumors (ie, islet cell tumors). The results of the association study showed that the mutation site (named T372R mutation) at position 115 (Chromosome 100, 743, 807 C-G) in SEQ ID NO: 1 was in the control group and the case group. There is a significant difference in distribution (0.05), so it can be used as a specific genetic locus (SNV) for the detection of auxiliary diagnosis. The present invention has been completed on this basis.
  • the inventors found a high frequency T372R mutation in the YY1 (Yin Yang 1) gene, and 113 functional pancreatic nerves. This site was found to be highly mutated in all tumors by sequencing in endocrine tumors at 30% (34/1 13).
  • the hotspot mutation of this gene reveals for the first time the important role of the T372R site of YY1 gene in the development of pancreatic neuroendocrine tumors, and can be used as a diagnostic marker and drug target for pancreatic neuroendocrine tumors, ie, the YY1 gene with this site. Mutations are more suitable for treatment with mTOR (eg, everolimus).
  • the sequence of the YY1 gene is known, and its detailed sequence and related information can be found at http://www.ncb.im.nih.gov/Genebank/.
  • the nucleotide sequence of the YY1 gene associated with the present invention is given in SEQ ID NO: 1.
  • the ⁇ gene is located on chromosome 14 and encodes the YY1 protein.
  • the YY1 protein is a widely expressed and conserved transcription factor that plays an important role in metabolic regulation, multiple tumorigenesis, and epigenetic regulation. Pancreatic neuroendocrine tumor-associated mutation
  • the present inventors have examined the results of genome-wide association and validation studies, and the results indicate that the mutation at position 1 115 in SEQ ID NO: 1 is a site highly correlated with pancreatic neuroendocrine tumorigenesis.
  • This high frequency T372R mutation has a very high mutation frequency in all sporadic islet cell tumors, which is about 30% (34/113).
  • the present invention discloses the correlation between the T372R site mutation of the YY1 gene and the occurrence of pancreatic neuroendocrine tumors.
  • the term "gene mutation site of the invention” or “inventive SNV” refers to the 1115th C/G mutation in the nucleotide sequence of the YY1 gene (shown as SEQ ID NO.: 1), etc.
  • SEQ ID NO.: 1 the nucleotide sequence of the YY1 gene (shown as SEQ ID NO.: 1), etc.
  • the frequency of genotype G in pancreatic neuroendocrine tumors was significantly higher than in normal controls. No such mutation site is present in the normal control DNA sequence.
  • the YY1 gene or the corresponding nucleic acid molecule or polypeptide molecule has various aspects. New use. These uses include (but are not limited to):
  • pancreatic neuroendocrine tumors such as whether it is suitable for the treatment of everolimus
  • An agent or kit for the preparation of a pancreatic neuroendocrine tumor for the auxiliary diagnosis a kit for preparing a patient for judging a pancreatic neuroendocrine tumor suitable for treatment with everolimus. Detection method, detection reagent and kit
  • Mutation sites associated with the YY1 gene can be used for the auxiliary diagnostic typing of pancreatic neuroendocrine tumors, especially early adjuvant diagnosis.
  • the detection methods of the invention can be used to assess disease prognosis and treatment regimens in individuals with pancreatic neuroendocrine tumors.
  • the test sample used in the present invention is not particularly limited, and for detecting a mutation site, it may be DNA or mRNA extracted from a sample such as a cell or a tissue. Since the T372R mutation of the present invention is a somatic mutation, the mutation is present in islet cells or islet tumor cells, usually not in peripheral blood cells. Therefore, a preferred test sample is islet tissue or islet tumor cells.
  • a part or all of the gene sequence detection of the present invention can be immobilized as a probe on a microarray or a DNA chip (also referred to as a "gene chip” or a “nucleic acid chip”) for analyzing the sequence of genes in tissues and Differential expression analysis, as well as genetic diagnosis.
  • the corresponding transcripts can also be detected by RNA-polymerase chain reaction (RT-PCR) in vitro amplification using primers specific for the YY1 gene.
  • Detection can be directed to cDNA as well as to genomic DNA. Mutated forms of the YY1 gene include point mutations, translocations, deletions, recombinations, and any other abnormalities compared to normal wild-type DNA sequences. Mutations can be detected using existing techniques such as Southern blotting, DNA sequence analysis, PCR and in situ hybridization. In addition, mutations may affect the expression of related proteins, so Northern blotting and Western blotting can be used to indirectly determine whether a gene has a mutation.
  • the most convenient method for detecting the mutation site of the present invention is to obtain an amplification product by amplifying the YY1 gene of the sample with a YY1 gene-specific primer; and then detecting whether the single nucleotide mutation of the present invention (SNV) is present in the amplification product. ).
  • the primers are 15-50 bp in length, preferably 20-30 bp in length. Although it is preferred that the primer is fully complementary to the template sequence, it will be appreciated by those skilled in the art that in the presence of a certain non-complementary primer (especially the 5' end of the primer), the primer can also be specifically amplified (ie only Amplify the desired fragment).
  • Kits containing these primers and methods of using the same are within the scope of the present invention as long as the amplified product amplified by the primer contains the corresponding position of the gene mutation site of the present invention.
  • a preferred primer pair has the sequence of position 1 115 in SEQ ID NO: 1.
  • the length of the amplification product is not particularly limited, the length of the amplification product is usually from 100 to 2,000 bp, preferably from 150 to 1,500 bp, more preferably from 200 to 1,000 bp. These amplification products should contain the 11th 15th position in SEQ ID NO: 1.
  • the main advantages of the present invention include - the first experimental confirmation of a new somatic mutation and a close correlation with pancreatic neuroendocrine tumors, thereby providing a method and kit for the auxiliary diagnosis of pancreatic neuroendocrine tumors.
  • the invention can provide an extremely valuable auxiliary reference index for clinical diagnosis (especially early diagnosis) of pancreatic neuroendocrine tumors, thereby facilitating early diagnosis, typing diagnosis and early prevention of pancreatic neuroendocrine tumors.
  • the mutation site of the present invention Since the mutation site of the present invention has a very high correlation with pancreatic neuroendocrine tumors, it can be used not only for early auxiliary diagnosis of pancreatic neuroendocrine tumors, but also for some carriers to take reasonable measures before the onset of disease. Preventive measures, thereby improving the carrier's survival and quality of life, and therefore have extremely significant application value and social benefits. Of course, the final diagnosis should also be confirmed by routine testing methods.
  • the invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention.
  • sequence, protein structure and other useful information about each gene can be found in the following electronic database information -
  • Criteria for the diagnosis of functional pancreatic neuroendocrine tumors clinical symptoms such as hypoglycemia, assessment of blood insulin and blood glucose levels (prolonged oral glucose tolerance test), CT/PET-CT impact diagnosis, and pathological diagnosis of surgical removal of tumors.
  • Genomic DNA was extracted from liquid nitrogen-preserved islet cell tumors and matched peripheral blood samples, and formalin-fixed paraffin-embedded (FFPE) insulinoma tissue samples. DNA was prepared using QIAGEN's DNeasy kit and QIAamp DNA FFPE tissue kit.
  • Dewaxing is performed on FFPE organizations using standard techniques. Immunofluorescence staining of the samples was performed according to standard experimental procedures. The following major antibodies were used: polyclonal rabbit anti-YY1 antibody (1:100; OriGene, clone EPR4651) and polyclonal guinea pig anti-insulin (1:400, Dako). Secondary antibodies for immunofluorescence staining were purchased from Invitrogen and Dako. Images were taken using the Olympus microscope system.
  • Genomic DNA is derived from islet cell tumor tissue and its matched peripheral blood. Then use the adapter to connect the ends of the clip. Then, the extracted DNA was amplified and purified by PCR (LM-PCR), hybridized on a NimbleGenEZ44M human exome sequencing chip, and then washed to remove the non-hybridized fragment. The non-captured and captured LM-PCR products were assessed for enrichment by performing quantitative PCR. Each library captured was subjected to high-throughput sequencing on the Hiseq2000 platform and ensured that each sample met the required average sequencing coverage depth. The sequenced image files are read by Illumina's software (default parameters).
  • PCR was performed using a dual 96 ⁇ L GeneAmp PCR System 9700 (Appl i ed Bi osystems), using 20 ng of template DNA per reaction in each sample. Sequencing was performed using a 3730x1 DNA Analyzer (Appl ied Bi osystems). All sequences are analyzed by the serial analysis software (Appl i ed Bi osystems, version 5. 2). The following primers were used: 5'-CACCCAGGGCAGGAATG-3' human YY1 -F 1 (SEQ ID NO.: 3); 5' -CCTGTCTCCGGTATGGA-3' human YY1-R1 (SEQ ID NO.: 4).
  • RT-PCR real-time reverse transcription polymerase chain reaction
  • somatic mutations in 78 tumors including 21 nonsense mutations, 49 missense mutations, 1 termination mutation, 3 splice sites and 4 frameshift mutations, with an average of 8 in each tumor. (2-18) somatic mutations.
  • the inventors have found a recurring YY1 gene c. C11 15G/P. T372 site mutation. by
  • YY1 is the target of mTOR inhibitors.
  • RAD001 a novel mTOR inhibitor
  • the present invention provides a drug target for the action of everolimus in the islet cell tumor, and therefore based on the present invention, the genetic diagnosis of YY1 mutation in islet cell tumors is a clinical therapeutic strategy for such tumors, drugs Treatment options and response rate judgments are important.
  • the nucleotide sequence of the YY1 gene is present: that is, the mutation of the 1st 15th C ⁇ G in SEQ ID NO.: 1 is closely related to pancreatic neuroendocrine tumor disease. Therefore, based on this mutation, the YY1 gene-specific primer can be designed to be amplified by using the patient's DNA as a template.
  • a small amount of islet tissue samples of the test subject in the test group were obtained, and DNA was extracted using a conventional method.
  • the PCR primers in the islet tumor cell test kit were diluted to 1 ⁇ mol/ ⁇ 1, and the extracted DNA was used as a template to carry out a PCR reaction with the provided primers.
  • sequencing was performed using a ⁇ 3730 DNA sequencer, and sequence interpretation and SNV confirmation were performed using Polyphred software.
  • the C ⁇ G mutation at position 100, 743, and 807 of chromosome 14 can also be detected by pyrosequencing or mass spectrometry of the amplified product and the normal control. Test results:
  • the test of the repeated examples was repeated except that 80 individuals (no known symptoms of islet cell tumor before detection) were randomly selected for detection.
  • the islet cell tissue sample to be detected is extracted, and the DNA is extracted from it using a conventional method (or using a specific kit).
  • the PCR primers in the test kit were diluted to ⁇ ⁇ ⁇ / ⁇ 1, and the extracted DNA was used as a template to carry out a PCR reaction with the provided primers.
  • the PCR products were purified and sequenced using a ⁇ 3730 DNA sequencer, and sequence interpretation and SNV confirmation were performed using Polyphred software.
  • Pancreatic neuroendocrine tumor samples were divided into YY1 gene mutation group and YY1 gene wild type group, and total RNA of tumor cells was extracted and reverse-transcribed into cDNA (Qiagen kit) using real-time quantitative PCR method (Invi trogen) The expression of the downstream gene of YY1 was detected.
  • the construction strategy is shown in Figure 5.
  • Construction method GFP fluorescent protein was expressed under the control of mouse YY1 promoter. When mating with Cre mice to remove the stop element in the middle of oxp, the mutant Yyl protein will be expressed under the control of the mouse YY1 promoter. Due to the high GC content, the mouse YY1 promoter needs to be taken from the BAC by homologous recombination.
  • the mouse YY1 gene cDNA sequence is shown as SEQ ID NO.: 22 (atggcctcgggcgacaccctctacatcgccacggacggctcggagatgccggccgagatcgtggag ctgc
  • the primary cells and cell lines obtained from the transgenic mice of Example 5 were cultured using a conventional method.
  • the plasmid was transfected with MIN6 cells.
  • Use liposome 2000 (Invi trogen).
  • Human full-length YY1 gene cDNA was inserted into the pCMV6 vector to construct an expression plasmid.
  • the T372R mutation was introduced into the plasmid using the Qui kChange II site-directed mutagenesis kit (Stratagene).
  • mTOR inhibitor The rapamyc in was added to the cell culture medium at different experimental concentrations. Rapamycin and everolimus are structural analogs.
  • Ki67 in the islet cell proliferation marker of YY1 transgenic mice was significantly increased, and the increase in Ki67 was significantly inhibited after administration of mTOR inhibitor (Fig. 7).

Abstract

L'invention concerne des loci de gène de sensibilité tumorale pancréatique neuroendocrinienne et des procédés et des kits de détection. En particulier, l'invention concerne des loci de gène de sensibilité tumorale pancréatique neuroendocrinienne basés sur des résultats de séquençage du génome complet, les loci étant une mutation somatique T372R de haute fréquence d'un gène YY1 (Yin Yang1). L'invention concerne également des procédés de détection de la sensibilité tumorale pancréatique neuroendocrinienne et de classification du sous-type de la tumeur pancréatique neuroendocrinienne et des kits de détection correspondants.
PCT/CN2014/078833 2013-05-29 2014-05-29 Loci de gène de sensibilité tumorale pancréatique neuroendocrinienne et procédés et kits de détection WO2014190927A1 (fr)

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CN109486952A (zh) * 2018-12-20 2019-03-19 中国医学科学院北京协和医院 miR-495在制备胰腺神经内分泌肿瘤诊断工具中的应用

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