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Novel application of cucurbitacin B

A technology of cucurbitacin and signaling pathway, applied in the new application field of cucurbitacin B, can solve the problems of less research on signaling pathway and unclear mechanism of action

Inactive Publication Date: 2018-11-13
HUBEI UNIVERSITY OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the specific mechanism of action of cucurbitacin B is not clear at present. The research on cucurbitacin B mainly focuses on signaling pathways such as STAT3 and MAPK, and there are few studies on other signaling pathways.

Method used

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  • Novel application of cucurbitacin B
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  • Novel application of cucurbitacin B

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Cucurbitacin B up-regulates the expression of LATS1 in human colon cancer cells

[0035] 1. CuB treatment of two cancer cells

[0036] ① Divide SW620 cells into four groups, put them into L-15 medium with CuB concentrations of 0 μmol / L, 0.05 μmol / L, 0.1 μmol / L and 0.15 μmol / L and each containing 10% FBS, and store them at 37°C After culturing for 24 hours, SW620 cells treated with 0 μmol / L CuB, SW620 cells treated with 0.05 μmol / L CuB, SW620 cells treated with 0.1 μmol / L CuB, and SW620 cells treated with 0.15 μmol / L CuB were collected respectively.

[0037]② Divide HT29 cells into four groups, put them into DMEM medium with CuB concentrations of 0 μmol / L, 0.1 μmol / L, 0.2 μmol / L and 0.3 μmol / L and each containing 10% FBS, and culture them at 37°C for 24 Hours, 0 μmol / L CuB-treated HT29 cells, 0.1 μmol / L CuB-treated HT29 cells, 0.2 μmol / L CuB-treated HT29 cells, and 0.3 μmol / L CuB-treated HT29 cells were collected respectively.

[0038] 2. Detection

[0039] ...

Embodiment 2

[0042] Example 2 Cucurbitacin B down-regulates the expression of YAP and its downstream target genes in human colon cancer cells

[0043] Cell processing and detection are the same as in Example 1, such as figure 2 As shown, the results of Western Blot showed that the phosphorylation level of YAP increased with the increase of CuB concentration, and the protein levels of YAP and YAP downstream target genes decreased with the increase of CuB concentration, indicating that CuB can down-regulate Protein levels of YAP and YAP downstream target genes.

Embodiment 3

[0044] Example 3 Cucurbitacin B inhibits the growth of colon cancer cells

[0045] The colon cancer SW620 and HT29 cell lines in the logarithmic growth phase were inoculated in 96-well plates (approximately 5000 cells per well, 90 μl medium), and cultured (37 ° C, 5% CO 2 Incubator) 24 hours later, the two kinds of cells were treated with cucurbitacin B (Cucurbitacin B, CuB) with different concentration gradients (0.05-1 μmol / L). After culturing for 20 hours and 44 hours, 10 μl of 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) solution with a concentration of 5 mg / ml was added to each well, The incubation was continued for 4 hours. After the reaction was terminated, the culture medium was sucked off, 150 μl dimethyl sulfoxide was added to each well, shaken at a low speed to fully dissolve, and then the absorbance at 490 nm (OD490) was measured with a microplate reader to calculate the survival rate.

[0046] Experimental results refer to image 3 , the res...

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Abstract

The invention relates to novel application of cucurbitacin B. Particularly, studies find that the cucurbitacin B can upregulate expression of an LATS1 gene and inhibit expression of a YAP gene and downstream related genes thereof in a colon cancer cell line. Moreover, the cucurbitacin B has the effect of inhibiting proliferation, invasion and metastasis of a human colon cancer SW620 cell line andan HT29 cell line, and can promote apoptosis thereof. The invention finds that the cucurbitacin B can be used as an activator for regulating a Hippo-YAP signaling pathway, is a novel promising effective anticancer medicine, can be used as an active component in preparation of medicines for treating cancers such as human colon cancer, and is conducive to improving the targeting ability of medicinalcancer treatment and reducing toxic and side effects of the medicines.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a new application of cucurbitacin B. Background technique [0002] The Hippo signaling pathway is a signaling pathway that regulates organ size discovered through chimera screening in Drosophila. It is a kinase chain composed of a series of protein kinases and transcription factors. From lower animals to higher animals, the Hippo signaling pathway is highly conserved. The pathway mainly includes three parts: upstream regulatory elements (NF2, Mer, etc.), core components (mainly Mst1 / 2, LATS1 / 2) and downstream effector molecules (YAP). The Hippo signaling pathway regulates cell proliferation, apoptosis, and self-renewal of stem cells, and thus plays an important role in controlling organ size. In the Hippo signaling pathway, Mst1 / 2 kinase phosphorylates LATS1 / 2 kinase with the assistance of its scaffold protein Sav1, and phosphorylated LATS1 / 2 is further activated in the interaction w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/575A61P35/00
CPCA61K31/575A61P35/00
Inventor 司渊刘莹伍叶子许佳欣向雨晨张特
Owner HUBEI UNIVERSITY OF MEDICINE
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