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Haplotype of tumor metastatic suppressor gene HTPAP as well as construction method and application thereof

A technique for inhibiting gene and tumor metastasis, applied in the field of genetic engineering, can solve the problems of limited functions, insufficient comprehensiveness, and less information

Inactive Publication Date: 2009-09-30
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the function of a single SNP itself is limited, it is not comprehensive enough to evaluate the function of the HTPAP gene by using a single site SNP analysis, and it provides less information

Method used

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  • Haplotype of tumor metastatic suppressor gene HTPAP as well as construction method and application thereof
  • Haplotype of tumor metastatic suppressor gene HTPAP as well as construction method and application thereof
  • Haplotype of tumor metastatic suppressor gene HTPAP as well as construction method and application thereof

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Embodiment 1

[0088] 1. Laser capture microdissection (LCM) for cutting pure tumor cells and extracting DNA: take fresh tumor specimens, after O.C.T embedding, perform 8 μm serial frozen sections, and stick them on the coating specially used for LCM On the sheet, 10 sheets each. After rapid hematoxylin-eosin (HE) staining, turn on the Veritas laser capture microdissection system (Arcturus, USA), and cut pure tumor cells into the CapSure cap (be careful not to cut mixed stroma components). Pure tumor cell DNA was extracted using the QIAamp DNA Micro Kit (Qiagen, Germany) protocol, and the extracted DNA was measured by an ultraviolet spectrophotometer and stored at -20°C for SNP detection. All cases underwent LCM successfully, and enough pure tumor cell DNA was obtained ( figure 1 ).

[0089] 2. SNP detection: Pyrosequencing method can be used to quickly and accurately determine a short target fragment. The specific steps are as follows:

[0090] Step 1: A specific sequencing primer is co...

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Abstract

The invention relates to a haplotype of a tumor metastatic suppressor gene HTPAP as well as a construction method and an application thereof, belonging to the field of gene engineering. Through the SNP examination and the analysis of a patient with liver cancer, the invention constructs the haplotype of the HTPAP with the following structure: C-A-T-C-A, T-G-G-G-G, C-G-T-C-A, T-A-G-G-G, C-A-G-C-A, T-A-G-G-A, C-A-T-G-A, T-A-T-C-A, T-A-G-C-G, C-A-T-G-G or T-G-T-C-G. The haplotype can provide a guide for predicting the metastasis and the recurrence after a liver cancer operation, is simple, convenient and practical and can be used for evaluating the patient with liver cancer after prognosis and taking an integrated method to intervene in time so as to improve the long-dated survival rate of the patient, and providing the reference for improving the prognosis; and the haplotype can be combined with other molecular indexes, , so as to provide a new trial box for the diagnoses of the liver cancer recurrence and the prognosis prediction to be convenient for clinical application.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a haplotype of a tumor suppressor gene HTPAP and its construction method and application. Background technique [0002] The prior art discloses that deletion of chromosome 8p is related to the recurrence and metastasis of hepatocellular carcinoma (hereafter referred to as HCC), and a full-length gene HTPAP was cloned on the basis of this. The gene is a candidate liver cancer metastasis suppressor gene, and its official name is PPAPDC1B (Phosphatidic acid phosphatase type 2 domain containing1B), which is a type 2 phosphatidic acid phosphatase containing 1B functional domain. The gene is located at 8p12 and contains 7 exons and 6 introns. The full length of the gene is 4459bp, and the cDNA length is 826bp, encoding a protein with 175 amino acids. The homologous sequence of this gene exists widely in animals (such as chimpanzees, dogs, musculus musculus, etc.) and pl...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12Q1/68
Inventor 任宁钦伦秀汤钊猷叶青海武金才贾户亮孙海晶周海军
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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