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Application of HNF4 (hepatocyte nuclear factor 4) alpha proteins to preparation of liver cancer prognosis evaluation kit

A liver cancer prognosis, α protein technology, applied in the biological field, can solve problems such as prognosis judgment of liver cancer patients without HNF4α

Active Publication Date: 2013-12-11
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no research report that HNF4α can regulate the metastasis of liver cancer and be used to judge the prognosis of liver cancer patients

Method used

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  • Application of HNF4 (hepatocyte nuclear factor 4) alpha proteins to preparation of liver cancer prognosis evaluation kit
  • Application of HNF4 (hepatocyte nuclear factor 4) alpha proteins to preparation of liver cancer prognosis evaluation kit
  • Application of HNF4 (hepatocyte nuclear factor 4) alpha proteins to preparation of liver cancer prognosis evaluation kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 2×10 4 SMMC-7721 liver cancer cells overexpressing HNF4α and control cells overexpressing GFP were respectively inoculated on the upper layer of polycarbonate membrane chambers (products of Corning Company), and after 24 hours of culture, the cells passing through the polycarbonate membrane were stained with crystal violet. , observed and photographed under a microscope, counted the number of liver cancer cells passing through the polycarbonate membrane in each field of view, calculated the average value and calculated the standard deviation. The results showed that, compared with the control cells, the migration ability of liver cancer cells with high expression of HNF4α was significantly decreased in vitro ( figure 1 ), suggesting that HNF4α may inhibit liver cancer cell metastasis.

Embodiment 2

[0051] Forty-four 8-week-old nude mice were injected with 2×10 6 After stably expressing luciferase, SMMC-7721 liver cancer cells were divided into experimental group and control group (22 in each group). The experimental group was injected with adenovirus overexpressing HNF4α through the tail vein every week, and the mice in the control group were injected with adenovirus overexpressing GFP for 6 consecutive weeks. About 11 weeks after the cells were inoculated, the mice were intraperitoneally injected with the luciferase substrate, and the liver cancer metastasis was observed under an in vivo imager. The lungs of the sacrificed mice were observed and photographed, the number of metastases was counted and the volume was measured ( figure 2 ). The results showed that HNF4α could significantly inhibit the metastasis of liver cancer cells in vivo.

Embodiment 3

[0053] Randomly select paraffin sections of postoperative liver cancer tissues from 429 patients with liver cancer (the liver cancer tissue sections were all from Dongfang Hepatobiliary Surgery Hospital, and were diagnosed as hepatocellular carcinoma by 2 pathologists), and immunohistochemical methods were used to detect the expression of HNF4α protein in liver cancer tissues. Expression level and calculation of immunohistochemical score, the specific steps are as follows:

[0054] (1) Paraffin sections of liver cancer tissues were prepared, and the oven was set at 60° C. overnight;

[0055] (2) slices are dewaxed to water;

[0056] (Xylene Ⅰ①10min→XyleneⅡ②10min→XyleneⅢ③10min→100% ethanol 5min→95% ethanol 5min→85% ethanol 5min→75% ethanol 5min→double distilled water 5min)

[0057] (3) 3%H 2 o 2 Solution, placed at room temperature for 20min;

[0058] (4) Wash with double distilled water for 5min×3;

[0059] (5) Antigen retrieval: put slices into 0.01M citrate buffer (pH6....

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Abstract

The invention belongs to the technical field of biology. According to the invention, the protein expression index of HNF4 (hepatocyte nuclear factor 4) alpha proteins in liver cancer tissues is detected by an immunohistochemical method, so that the distant metastasis risk and the postoperative condition of a liver cancer patient can be judged. The invention provides an application of the HNF4 alpha proteins to preparation of a liver cancer prognosis evaluation kit. According to the application, the HNF4 alpha proteins are used as protein molecular markers for judging the prognosis condition of the liver cancer patient, so that the application has a guiding significance on postoperative monitoring and sequential treatment of the liver cancer patient.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the application of HNF4α protein, in particular to the application of HNF4α protein in the preparation of liver cancer prognosis assessment kit. Background technique [0002] Liver cancer is one of the most common malignant tumors, and its morbidity and mortality have remained high for a long time. my country is the country with the most incidence of liver cancer in the world, accounting for about 50% of the world's new cases each year. Statistics show that the high recurrence and metastasis rate after liver cancer surgery (5-year recurrence rate reaches 70%, small liver cancer also reaches 50%) has become a bottleneck for further improving long-term efficacy, and there are few indicators for judging the prognosis of liver cancer. [0003] Liver cancer metastasis is a complex process involving and interacting with multiple factors, which is not only related to the cancer cells themselv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 宁北芳谢渭芬丁劲张新尹川许文萍
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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