Molecular markers of early liver cancer and application of makers

A molecular marker and early liver cancer technology, applied in the field of early liver cancer molecular markers, can solve the problems of limited sensitivity and specificity, poor diagnostic effect of eHCC, etc., and achieve high detection sensitivity and specificity

Inactive Publication Date: 2011-09-07
JIANGSU ETERN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

More importantly, the sensitivity and specificity of the above-mentioned methods, whether molecular markers or imaging methods, are limited.

Method used

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  • Molecular markers of early liver cancer and application of makers
  • Molecular markers of early liver cancer and application of makers
  • Molecular markers of early liver cancer and application of makers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Collection of samples and protein extraction:

[0017] 1. Take human cirrhotic liver, atypical hyperplasia nodular lesions and early liver cancer lesions, and after microscopic diagnosis of HE slices by 2 pathologists, cut the corresponding wax blocks into white slices with a thickness of 7 μm. Dry at 60°C until there are no air bubbles between the sliced ​​tissue and the glass slide (about 30-60 minutes).

[0018] 2. Formaldehyde-fixed / paraffin-embedded (FFPE) tissue protein extraction and protein concentration determination:

[0019] 2.1. Dewax the above-mentioned liver tissue sections in xylene for 2 times, 10 min each time. After air-drying, the range of the lesion is accurately drawn. Use a scalpel blade (No. 11 blade) to cut the lesion precisely and put it into a low adsorption tube.

[0020] 2.2. Add an appropriate amount of paraffin-embedded tissue protein extraction buffer (Tris-HCl buffer) to each low adsorption tube, which contains 2% sodium dod...

Embodiment 2

[0023] Example 2: Western blotting demonstrated the differences in the expression of the three proteins in liver cirrhosis, dysplastic nodules and early liver cancer:

[0024] 1. The protein samples extracted in Example 1 (mixed with 18 samples in each group) were subjected to sodium dodecylsulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) by conventional methods, and then transferred to PVDF membrane (Millipore) on.

[0025] 2. Dissolve skim milk powder in TBS-T buffer (0.05% (v / v) Tween 20 / Tris-buffer) at pH=7.4 to obtain fat-free milk solution, the concentration of skim milk powder is 5% (w / v ), add the above fat-free milk solution to the above protein sample, add the primary antibody after 1h, incubate overnight at 4°C, wash the membrane with TBS-T for 3 times, add horseradish peroxidase (HRP)-labeled secondary Antibody, incubated at room temperature for 1h. Among them, the company and dilution ratio of the primary antibody are as follows: mouse anti-ACY1 (ab54960,...

Embodiment 3

[0027] Embodiment 3: Immunohistochemical method proves the expression difference of 3 kinds of proteins:

[0028] 1. Take human cirrhotic liver, atypical hyperplasia nodular lesions and early liver cancer lesions. After microscopic diagnosis of HE slices by 2 pathologists, the corresponding wax blocks are made into tissue chips by conventional methods and cut into 4μm thickness. Sections were placed on slides coated with 3-aminopropyltriethoxysilane.

[0029] 2. Dewax the slides with tissue sections according to conventional methods, and then hydrate them with 100% ethanol, 95% ethanol, and 85% ethanol for 5 minutes each. Place the slides with tissue sections in In 0.01M citrate buffer (pH6.0), carry out antigen retrieval treatment at 100°C for 3 minutes, take it out and let it cool down, and place it in H2O with a mass concentration of 3%. 2 o 2 Endogenous peroxidase was blocked in phosphate (PBS, pH=7.4) buffer solution for 30 minutes, removed, placed in goat serum for 60 ...

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Abstract

The invention provides molecular markers of early liver cancer and application of the markers. The molecular markers of early liver cancer comprise amino-acylase-1 (ACY1), sulphite oxidase (SUOX) and glycogen phosphorylase (PYGL). The invention further provides an application of ACY1, SUOX and PYGL as molecular markers of early liver cancer in preparation of liver cancer clinical diagnostic reagents. The invention further provides a liver cancer diagnostic kit, wherein the liver cancer diagnostic kit is characterized by comprising ACY1 antibody, SUOX antibody and PYGL antibody. In the invention, new molecular markers of early liver cancer are discovered and have relatively high detection sensitivity and specificity.

Description

technical field [0001] The invention relates to a group of early liver cancer molecular markers and applications thereof. Background technique [0002] The formation of liver cancer is a complex process involving multiple factors, multiple steps, and multiple genes. It generally goes through the progression of hepatitis, liver cirrhosis, dysplasia, and liver cancer. In recent years, dysplastic nodules (DN) have been paid more and more attention as precancerous lesions of liver cancer. DN in cirrhotic liver is widely recognized as a precancerous lesion of liver cancer. At present, the clinical differential diagnosis of dysplastic nodules and early liver cancer (according to BCLC regulations, eHCC is defined as a single tumor or the sum of three tumors <3 cm in size, Child-Pugh A and B, and does not meet the criteria for liver transplantation) mainly relies on There is still a lack of effective immunohistological diagnostic markers for imaging diagnosis or microscopic dia...

Claims

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Application Information

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IPC IPC(8): G01N33/574
Inventor 金光植刘银坤王浙东
Owner JIANGSU ETERN
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