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Application of vitronetin

A technology of vitronectin and protein, applied in the direction of instruments, measuring devices, scientific instruments, etc.

Inactive Publication Date: 2007-07-25
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] But so far, there is no report about the direct correlation between Vitronectin and hepatocellular carcinoma and its mechanism of action in the prior art

Method used

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  • Application of vitronetin
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 , Preparation of protein samples from hepatocellular carcinoma tissue and paracancerous tissue

[0026] Urea, 3-[(3-cholamidopropyl)-diethylammonium]-1-propanesulfonic acid (CHAPS), phenylmethylsulfonyl fluoride (PMSF), dithiothreitol used in this example (DTT) were purchased from Sigma Company.

[0027] In this example, protein samples of cancer tissue and paracancerous tissue of hepatocellular carcinoma were prepared by nonenzymatic sample preparation (NESP), as follows:

[0028] 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

[0033] Example 2 , Screening of differentially expressed proteins

[0034] Urea, 3-[(3-cholamidopropyl)-diethylammonium]-1-propanesulfonic acid (CHAPS), sodium dodecylsulfonate (SDS), dithiothreitol used in this example (DTT) was purchased from Sigma; iodoacetamide (IAA), acrylamide, N, N-methylenebisacrylamide, etc. were purchased from Fluka; cleavable ICAT reagents were purchased from Applied Biosystems Framingham, MA.

[0035] Ammonium persulfate (AP), TEMED, Tri-n-butylphosphat (TBP), PDQuest software, etc. are Bio-Rad products.

[0036] Avidin affinity column was purchased from Applied Biosystems, Framingham, MA.

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

[0038] The composition of the loading buffer used is: 0.6ml of 1mol / L Tris-HCl (pH6.8), 5ml of 50% glycerol, 2ml of 10% SDS, 0.5ml of mercaptoethanol, 1.9ml of distilled water, and a small amount of hexabromophenol blue.

[0039] First, the gel-enhan...

Embodiment 3

[0047] Example 3 , Vitronectin differential expression of Western blot verification

[0048] In order to confirm the differential expression of Vitronectin, 10 hepatocellular carcinoma patients' cancer tissues and corresponding paracancerous tissue protein samples (prepared by NESP method, other 10 cases except 429 cases in Table 1) were taken, and the purchased anti-cancer Vitronectin antibody was used for western blot analysis, the specific process is briefly described as follows:

[0049] 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 vitronectin monoclonal antibody (purchased from Abcam Ltd, 1:200 dilution) ), incubated at room temperature for 2 hours, washed three times with TBST (containing 2.42g of Tris, 8g of sodium chloride per liter, 201ml of Tween, and adjusted the pH to 7.6 with HCl), 5 minutes each time, and the se...

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Abstract

An application of Vitronectin includes finding out a high expression protein in cancer tissue of hepatocyte cancer by selecting differential expression protein in cancer tissue and cancer side tissue of hepatocyte cancer, confirming that differential expression of Vitronectin is definitely existed in cancer tissue and side cancer tissue of hepatocyte cancer by immune print test, enabling to use Vitronectin as protein molecule label to detect hepatocyte cancer based on relativity of Vitronectin to hepatocyte cancer.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to the application of Vitronectin as a protein molecular marker for detecting 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 identify new Liver cancer-related...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/577
Inventor 曾嵘李辰周晓袁新雨
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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