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Application of substance taking MANF as target spot in preparing products for treating intrahepatic cholangiocarcinoma

A technology for intrahepatic cholangiocarcinoma and intrahepatic cholangiocarcinoma, applied in the application field of substances targeting MANF in the preparation of products for the treatment of intrahepatic cholangiocarcinoma, which can solve the difficulties of early diagnosis, low radical resection rate, poor prognosis of ICC, etc. question

Active Publication Date: 2019-02-22
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 substance taking MANF as target spot in preparing products for treating intrahepatic cholangiocarcinoma
  • Application of substance taking MANF as target spot in preparing products for treating intrahepatic cholangiocarcinoma
  • Application of substance taking MANF as target spot in preparing products for treating intrahepatic cholangiocarcinoma

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Experimental program
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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 an application of a substance taking MANF as a target spot in preparing products for treating intrahepatic cholangiocarcinoma. An experiment proves that the substance for inhibiting MANF expression in cells or inhibiting MANF activity in cells is capable of effectively inhibiting the multiplication, migration and / or invasion of cholangiocarcinoma cells, is capable of inhibiting the growth of cholangiocarcinoma and can be used for treating or / and preventing 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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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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