Application of shizukaol D in preparing anti-tumor target medicines
An anti-tumor and drug technology, applied in the fields of medicine and genetic engineering, can solve the problem that there is no report of shizukaolD targeting and inhibiting cell signaling pathway.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0093] Example 1 The signal of Wnt pathway is inhibited
[0094] Take 293T cells and plant a 24-well plate (4~6×10 per well 4 Cells). After adherence, it was treated with 50mmol / L LiCl for 12 hours, and then transferred to Topflash, Fopflash and internal reference RL plasmids. Shizukaol D and DMSO control were added the next day (that is, 0.1% DMSO was added as a control) for 24 hours, and 3 replicates were made for each sample. Receive samples and detect luciferase activity. Take XAV939 as a positive control. XAV939 is a tankryase enzyme inhibitor that can indirectly inhibit the Wnt pathway (Huang SM, Mishina YM, Liu S, et al. Tankyrase inhibition stabilizes axin and antagonizes Wnt signalling[J].Nature,2009,461(7264):614-20. ). XAV-939 selectively inhibits Wnt / β-catenin-mediated transcription by inhibiting tankyrase1 / 2, IC 50 It is 11nM / 4nM, which regulates the level of axin, but has no effect on CRE, NF-κB and TGF-β.
[0095] Such as figure 1 As shown, shizukaol D at 6.25 a...
Embodiment 2
[0096] Example 2 Verification of differential genes in Wnt pathway
[0097] After SMMC-7721 cells adhere to the wall, shizukaol D is added, and samples are collected at 0, 6 and 12 hours respectively. RNA is extracted by Trizol method, and genomic RNA is removed. After reverse transcription, real-time PCR is used to detect gene expression in the samples. Use GAPDH as an internal reference. Real-time Quantity PCR (Q-PCR) was used to detect the difference in the number of amplification cycles (-ΔΔCt) of related genes in the Wnt pathway to determine the change in the expression of the gene.
[0098] Such as figure 2 As shown, the expression levels of c-Myc, cyclin D, Tcf-1, LEF1, wnt3a and FGF18 were all decreased in the samples with shizukaol D added for 12 hours compared with the samples without shizukaol D. QPCR results showed that shizukaol D down-regulated the expression of c-myc and other genes. This may be because shizukaol D can inhibit the expression of β-catenin, thereby ...
Embodiment 3
[0100] Example 3 The addition of wnt3a partially offsets the inhibitory effect of shizukaol D on the growth of liver cancer cells
[0101] Adding wnt3a is a classic way to activate the Wnt pathway. The medium of L-wnt3a cells contains sufficient wnt3a, which usually activates the Wnt pathway of the cells. Take SMMC-7721 cells, add medium containing wnt3a after adherence, and incubate overnight. After adding 6.25 and 12.50μmol / L shizukaol D, after 48 hours of treatment, use CCK-8 kit to detect cell viability. Take 0.1% DMSO as control.
[0102] The result is image 3 As shown, adding medium containing wnt3a can significantly improve the survival rate of SMMC-7721 cells under the action of shizukaol D. Regardless of whether 6.25 or 12.50μmol / L shizukaol D is added, once wnt3a is added to the medium, the viability of SMMC-7721 cells can be restored to the same level, which shows that adding wnt3a to the medium can offset the difference in compound concentration between the two grou...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com