Application of substances targeting manf in the preparation of products for the treatment of intrahepatic cholangiocarcinoma

A technology for intrahepatic cholangiocarcinoma and substances, which is applied in the field of application of substances targeting MANF in the preparation of products for the treatment of intrahepatic cholangiocarcinoma, which can solve the problem of poor prognosis of ICC, low rate of radical resection, and loss of optimal surgery for patients Timing and other issues

Active Publication Date: 2021-09-24
ANHUI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are two main reasons for the poor prognosis of ICC: (1) The clinical features of ICC are not obvious, early diagnosis is difficult, and lymph node and distant metastasis are prone to occur. Low and unstable; (2) The pathogenesis of ICC is unclear and there is a lack of effective targeted therapy drugs

Method used

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  • Application of substances targeting manf in the preparation of products for the treatment of intrahepatic cholangiocarcinoma
  • Application of substances targeting manf in the preparation of products for the treatment of intrahepatic cholangiocarcinoma
  • Application of substances targeting manf in the preparation of products for the treatment of intrahepatic cholangiocarcinoma

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Experimental program
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Effect test

preparation example Construction

[0052] 1) Preparation of protein samples:

[0053] For cultured cell lines, discard the culture supernatant, pipette with pre-cooled 1×PBS solution, collect the cells and wash 3 times, discard the PBS, add an appropriate volume of 1×RIPA lysate containing protease inhibitors, pipette and mix well After that, place it on ice for 30 minutes, and centrifuge at 12000 rpm for 10 minutes at 4°C. The supernatant was transferred to a new EP tube and set aside.

[0054] 2) Treatment of protein samples before electrophoresis:

[0055] Add 1 / 4 volume of 4×SDS-PAGE protein loading buffer to the protein lysate, boil at 100°C for 5-10min, cool to room temperature and load the sample.

[0056] 3) Prepare 12% SDS-PAGE gel, add sample, electrophoresis, transfer membrane, incubate primary antibody and secondary antibody, and expose for imaging. Among them, the primary antibody is a rabbit anti-MANF polyclonal antibody (ab23621, Abcam, USA), and the secondary antibody is a mouse secondary ant...

Embodiment 1

[0057] Example 1. The shRNA that targets human MANF and interferes with the expression of human MANF gene inhibits the proliferation, migration and / or invasion of cholangiocarcinoma cells, and inhibits the growth of cholangiocarcinoma tumors

[0058] 1.MANF is highly expressed in ICC

[0059] The inventors of the present application not only detected the expression level of human MANF in human hepatocellular carcinoma (HCC) tissue and its adjacent tissue by immunohistochemistry, but also detected the expression level of human MANF in human ICC tissue and its adjacent tissue. The expression level. The results showed that the level of human MANF in human hepatocellular carcinoma tissue was significantly lower than that in the paracancerous control. In human intrahepatic cholangiocarcinoma tissue, the expression of human MANF was detected by immunohistochemistry, and it was found that the level of human MANF was significantly higher than that of adjacent tissues ( figure 1 ). ...

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Abstract

The invention discloses the application of a substance targeting MANF in the preparation of a product for treating intrahepatic cholangiocarcinoma. Experiments have proved that the substances that inhibit the expression of MANF in cells or the substances that inhibit the activity of MANF in cells can effectively inhibit the proliferation, migration and / or invasion of cholangiocarcinoma cells, inhibit the growth of cholangiocarcinoma tumors, and can be used to treat intrahepatic cholangiocarcinoma or / and prevent Intrahepatic cholangiocarcinoma.

Description

technical field [0001] The invention relates to the application of MANF-targeted substances in the preparation of products for treating intrahepatic cholangiocarcinoma. Background technique [0002] Intrahepatic cholangiocarcinoma (ICC) is a malignant tumor originating from intrahepatic bile duct epithelial cells. It is the second most common malignant tumor of the liver after hepatocellular carcinoma, accounting for about 10%-15% of primary liver cancers. , and its incidence has been on the rise worldwide in recent years. Its 5-year survival rate is about 5%-15%, and the average survival period without surgery is only 9 months. There are two main reasons for the poor prognosis of ICC: (1) The clinical features of ICC are not obvious, early diagnosis is difficult, and lymph node and distant metastasis are prone to occur. Low and unstable; (2) The pathogenesis of ICC is unclear, and there is a lack of effective targeted therapy drugs. Therefore, studying the mechanism of I...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K31/7088A61P35/00A61P1/16
CPCA61K31/7088A61K45/00A61P1/16A61P35/00
Inventor 沈玉先李红张晓节吴正升
Owner ANHUI MEDICAL UNIV
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