Molecular markers of well-differentiated early liver cancer and use thereof

A molecular marker, early-stage liver cancer technology, applied in the field of molecular markers of well-differentiated early-stage liver cancer, can solve the problem of no difference in the expression of SQSTM1, and achieve the effect of high detection sensitivity and specificity

Inactive Publication Date: 2011-09-07
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has also been reported that it is increased in breast cancer and liver cancer, but there is no report of the diffe

Method used

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  • Molecular markers of well-differentiated early liver cancer and use thereof
  • Molecular markers of well-differentiated early liver cancer and use thereof
  • Molecular markers of well-differentiated early liver cancer and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0017] 1. Human atypical hyperplasia nodular lesions and early liver cancer lesions were diagnosed under the microscope by HE slices of 2 pathologists, and the corresponding wax blocks were cut 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 dodecylsulfon...

Embodiment 2

[0023] Example 2: Immunoblot method proves the expression difference of phosphoglucomutase-1 and sequestrum-1:

[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-PGM1 (ab55616, Abcam, 1:300), rabbit anti-SQSTM1 (P0067, Sigma-Aldrich,...

Embodiment 3

[0027] Example 3: Immunohistochemical method proves the expression difference of PGM1 and SQSTM1:

[0028] 1. After taking human atypical hyperplasia nodular lesions and early liver cancer lesions and diagnosing them under the microscope with the HE slices of two pathologists, the corresponding wax blocks were made into tissue chips by conventional methods, cut into 4 μm thick slices, and placed onto 3-aminopropyltriethoxysilane-coated glass slides.

[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) buffer solution for 30 minutes, removed, placed in goat serum for 60 minutes, ...

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Abstract

The invention provides a group of molecular markers of well-differentiated early liver cancer and a use thereof. The group of molecular markers of well-differentiated early liver cancer comprises phosphoglucomutase-1(PGM1) and sequestra sequestrum-1 (SQSTM1). The invention further provides a use of phosphoglucomutase-1 and sequestra sequestrum-1 as a molecular marker combination 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 a phosphoglucomutase-1 antibody and a sequestra sequestrum-1 antibody. The invention has the advantage that relatively high detection sensitivity and specificity are provided.

Description

technical field [0001] The invention relates to a group of molecular markers of well-differentiated early liver cancer and its application. Background technique [0002] Hepatitis is a social health problem that seriously affects human health in China and the world. Most patients with hepatitis are hepatitis B or hepatitis C, while hepatitis B is the main disease in my country. Although the incidence of hepatitis B in my country remains high, about 25% of chronic HBV-infected patients eventually develop hepatocellular carcinoma (HCC). 300 to 400 million people in the world are chronically infected with HBV, and there are more than 93 million patients with chronic HBV infection in China. At present, it has been proved that the hepatitis viruses related to liver cancer are mainly HBV and HCV. A long-term prospective study of hepatitis B surface antigen (HBsAg) carriers found that the relative risk (RR) of HBsAg-positive patients developing liver cancer was 13.69 times that o...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N33/573G01N33/48
Inventor 金光植刘银坤李岩康晓楠郭坤
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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