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

A technology for inhibiting gene and tumor metastasis, applied in the field of genetic engineering, can solve problems such as insufficient comprehensiveness, limited function, and low amount of information

Inactive Publication Date: 2012-09-26
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 (Laser Capture Microdissection, LCM) for cutting of pure tumor cells and extraction of DNA: take fresh tumor specimens, and after OCT embedding, make 8μm continuous frozen sections and paste them on the film dedicated to LCM On the film, 10 each. After rapid hematoxylin-eosin (HE) staining, turn on the Veritas laser capture microdissection system (Arcturus, USA), and cut out pure tumor cells in the CapSure cap (be careful not to cut out mixed interstitial components). Use QIAamp DNA Micro Kit (German Qiagen company) to extract pure tumor cell DNA. The extracted DNA is measured by UV spectrophotometer and stored at -20°C for SNP detection. LCM was successfully performed in all cases, and enough pure tumor cell DNA was extracted and obtained ( figure 1 ).

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

[0090] Step 1: Combine a ...

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

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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