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Application of bone morphogenetic protein 4 in cancer inhibition

A technology for inhibiting liver cancer and protein, applied in the field of biopharmaceuticals, can solve the problems of liver cancer CSC differentiation and its regulatory mechanism is not very clear.

Active Publication Date: 2013-12-25
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] But so far, researchers have not been able to understand the CD133 + The differentiation and regulation mechanism of hepatocellular carcinoma CSC is not very clear

Method used

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  • Application of bone morphogenetic protein 4 in cancer inhibition
  • Application of bone morphogenetic protein 4 in cancer inhibition
  • Application of bone morphogenetic protein 4 in cancer inhibition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0190] Exogenous high concentration of BMP4 can induce down-regulation of CD133 protein expression in liver cancer cells

[0191] Mammalian liver development is regulated by a variety of growth factors. In order to screen a large number of growth factors that can induce the differentiation of liver CSCs, the inventors first analyzed and detected different liver cancer cells by flow cytometry in this example. The positive rate of CD133 protein expression in the line ( figure 1 A), the PLC / PRF / 5 cell line with a higher positive rate was selected as the screening cell, and the CD133+ / - liver cancer cell group with a sorting purity greater than 95% was obtained by flow cytometry ( figure 1 B).

[0192] The inventors detected the sorted CD133+ liver cancer cells, and found that the CD133+ cell population in the PLC / PRF / 5 cell line had higher non-adherent growth and self-renewal abilities than the CD133- cell population, and its stem The expression level of sex-related genes is al...

Embodiment 2

[0196] BMP4-induced reduction of CD133 protein expression in liver cancer cells is concentration- and time-dependent

[0197] In order to further observe the effect of BMP4 on liver cancer cells, especially liver cancer CD133+CSCs, in this example, the inventors treated PLC / PRF / 5 cells with different concentrations of BMP4 within the range of 0-100 ng / ml.

[0198] The results of Western Blot showed that BMP4 treatment at a concentration above 30ng / ml for 6 days could effectively induce a decrease in the expression of CD133 protein in the CD133+ liver cancer cell population, and this expression down-regulation was the strongest in the 100ng / ml BMP4 treatment group, but at the BMP4 concentration When it reaches 50ng / ml, it can already cause enough obvious effects ( image 3 A). The inventors chose a BMP4 concentration of 50 ng / ml to carry out subsequent experiments. At the same time, different concentrations of exogenous BMP4 did not cause changes in the expression of CD133 pr...

Embodiment 3

[0203] Effects of exogenous BMP4 on the proliferation and death of liver cancer cells

[0204] In this example, BrdU incorporation experiments were used to detect the effects of adding 0-100 ng / ml of exogenous BMP4 on the proliferation of PLC / PRF / 5CD133+ / CD133- cells.

[0205] The results showed that 10ng / ml and 20ng / ml BMP4 could promote the proliferation of PLC / PRF / 5 cells, but 50ng / ml and 100ng / ml BMP4 could have a certain inhibitory effect on the proliferation of CD133+ liver cancer cells. The effect is similar to that in the CD133+ cell group ( Figure 4 A).

[0206] In order to observe the effect of BMP4 on the apoptosis of liver cancer cells, the apoptosis rate of PLC / PRF / 5 cells after BMP4 treatment for 3 days was detected by Annexin V / 7-AAD double labeling method.

[0207] The results showed that BMP4 treatment failed to significantly affect the apoptosis of liver cancer cells ( Figure 4 B).

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Abstract

The present invention relates to an application of bone morphogenetic protein 4 in cancer inhibition. Specifically bone morphogenetic protein (BMP) has a liver cancer stem cell differentiation induction effect, wherein CD133 protein expression of liver cancer cell lines can be down-regulated and liver cancer cell line CD133+ liver cancer stem cell differentiation can be promoted with BMP4, and BMP4 can further be provided for inhibiting in vitro proliferation and self-renewal of liver cancer cell lines and inhibiting tumorigenicity of immune-deficient mice. The present invention further provides BMP4-induced liver cancer stem cell differentiation action mechanism.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, in particular, the invention relates to the application of bone morphogenic protein 4 in inhibiting cancer. Background technique [0002] Hepatocellular carcinoma (HCC) is the fifth most common tumor worldwide and the third leading cause of cancer death. With the continuous development of tumor research, surgical treatment, radiotherapy and chemotherapy have successfully reduced the tumor volume, but postoperative recurrence and metastasis of liver cancer are still very common, often leading to the final death of liver cancer patients. In recent years, in response to the characteristics of tumors that are prone to recurrence and metastasis, researchers discovered cancer stem cells (Cancer Stem Cells, CSCs), and believed that the occurrence and development of cancer stem cells played an important role in liver cancer. [0003] Cancer stem cells (CSCs) are a group of special tumor cells that exis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K9/00A61K45/00A61P35/00A61P1/16
Inventor 李锦军张立行
Owner SHANGHAI INST OF ONCOLOGY
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