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Application of nucleoside diphosphate kinase A

A technology for nucleoside diphosphate kinase and liver cancer, which is applied in the field of detection of susceptibility to liver cancer, and can solve the problems of unseen nucleoside diphosphate kinase A correlation

Inactive Publication Date: 2007-08-01
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The protein is highly expressed in neuroblastoma tissue, but it is negatively correlated with the invasion and metastasis of the tumor
[0007] So far, although many studies have shown that nucleoside diphosphate kinase A is negatively correlated with the invasion and metastasis of many cancers, there is no direct report on the correlation between nucleoside diphosphate kinase A and liver cancer

Method used

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  • Application of nucleoside diphosphate kinase A
  • Application of nucleoside diphosphate kinase A
  • Application of nucleoside diphosphate kinase A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 , Preparation of protein samples from hepatocellular carcinoma tissue and paracancerous tissue

[0025] Urea, 3-[(3-cholamidopropyl)-diethylammonium]-1-propanesulfonic acid (CHAPS), phenylmethylsulfonyl fluoride (PMSF), dithiothreitol used in this example (DTT) were purchased from Sigma Company.

[0026]In this example, protein samples of cancer tissue and paracancerous tissue of hepatocellular carcinoma were prepared by nonenzymatic sample preparation (NESP), as follows:

[0027] The surgically excised fresh tissue block is quickly placed on ice and quickly cut into several small pieces that are visible to the naked eye without areas of necrosis. Use pre-cooled glutamine-free RPMI1640 medium (5% fetal bovine serum, 0.2mM PMSF, 1mM EDTA, benzisoxazole penicillin 25mg / mL, gentamicin 50mg / mL, penicillin 100U / mL, Streptomycin 100mg / mL, Amphotericin B 0.25mg / mL, Nystatin 50U / mL) After washing the small tissue pieces several times, they were quickly ground into ...

Embodiment 2

[0032] Example 2 , Screening of differentially expressed proteins

[0033] Urea, 3-[(3-cholamidopropyl)-diethylammonium]-1-propanesulfonic acid (CHAPS), L-glutamine, C 12 L-leucine, C 13 L-leucine, L-lysine, etc. were purchased from Sigma.

[0034] Calcium chloride and sodium bicarbonate were purchased from Shanghai Sangon Bioengineering Co., Ltd.; magnesium chloride and magnesium sulfide were purchased from Shanghai Zhenxin Reagent Factory.

[0035] Dialyzed calf serum was purchased from GIBCO Company; iodoacetamide (IAA) was purchased from Fluka Company; pancreatin was purchased from Promage Company.

[0036] Tris (TRIS), PDQuest software, etc. are Bio-Rad products.

[0037] LTQ TM Deca XP system and ProteomeX TM Workstation was purchased from Thermo Finnigan Company.

[0038] Normal liver cell line L-02 and liver cancer cell line BEL-7404 were purchased from the Cell Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

[0039] The no...

Embodiment 3

[0050] Example 3 , Nucleoside diphosphate kinase A differentially expressed protein Western blot verification

[0051] In order to confirm the differential expression of nucleoside diphosphate kinase A, the cancer tissues and corresponding paracancerous tissue protein samples of 10 hepatocellular carcinoma patients prepared in Example 1 were taken, and the purchased anti-nucleoside diphosphate kinase A antibody was used for western blot analysis , the specific process is briefly described as follows:

[0052] Take 20 μg of protein samples from each sample and use 12% SDS-PAGE to separate, transfer to PVDF membrane (purchased from Amersham Biosciences Company), the primary antibody uses mouse anti-human nucleoside diphosphate kinase A monoclonal antibody (purchased from BioGenex Company, 1: 100 dilution), incubated at room temperature for 2 hours, washed three times with TBST (containing 2.42g Tris, 8g sodium chloride, 20ml Tween per liter, adjusted pH to 7.6 with HCl), 5 min...

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Abstract

This invention relates to one method to filter liver cell cancer organism and side organism difference expression protein, which finds out one liver cell cancer organism expression protein, wherein, the immune experiment further proves G protein beta2 subunit in liver cell cancer organism different expression; The protein can be used as protein molecule label for liver cancer by the relative property.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to an application of nucleoside diphosphate kinase A as an indicator for detecting the susceptibility of liver cancer. Background technique [0002] Liver cancer is a disease that seriously endangers human beings. The incidence of liver cancer in western developed countries is low, and the basic research on liver cancer in the world is still relatively weak. However, my country is a country with a high incidence of liver cancer, and the morbidity and mortality are on the rise, and the age of onset is younger. Medical expenses have increased greatly, and liver cancer has become the number one enemy that seriously endangers the safety of people's life and property in our country, and is an important factor affecting social and economic development. It is of strategic significance to intensify the basic research of liver cancer in my country, and to isolate and identi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/577
Inventor 曾嵘周晓李荣霞袁新雨
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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