Application of zju-imb-z19 compound in drug for treating prostate cancer
A technology of ZJU-IMB-Z19, 1. ZJU-IMB-Z19, applied in the field of prostate cancer drugs, can solve problems such as upregulation and enzalutamide resistance
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0035] Example 1 ZJU-IMB-Z19 can inhibit the transcriptional function of endogenous and exogenous AR
[0036] will be 1.0 x 10 5 Cells / mL of PC-3 or LNCaP cell suspension were seeded in 24-well plates at 500 μL per well. When the cells grew to 60%, PC-3 cells were co-transfected with 100 ng PSA-luc, 20 ng AR-WT and 1 ng pCMV-Renilla plasmids per well (LNCaP cells were transfected with 100 ng PSA-luc and 1 ng pCMV-Renilla plasmids). 6 hours after transfection, the medium was changed to phenol red-free RPMI 1640 medium containing 10% charcoal-treated fetal bovine serum (FBS); 24 hours after transfection, one group was added with 5nM of dihydrotestosterone per well. The concentration of ZJU-IMB-Z19 or enzalutamide was added 1 μL each, and the other group was added 1 μL of DMSO (dimethyl sulfoxide) or the corresponding concentration of ZJU-IMB-Z19 or enzalutamide, and the culture was continued for 24 Hour. Finally, the medium was aspirated, and 100 μL of 1×PLB was added to each...
Embodiment 2
[0038] Example 2 ZJU-IMB-Z19 can inhibit the transcriptional activity of endogenous AR in LNCaP cells at the protein level without affecting the expression of endogenous AR.
[0039] Only detecting the inhibition of AR on downstream target genes by ZJU-IMB-Z19 by the dual-luciferase reporter system cannot fully explain the antagonistic effect of this compound on AR. The present invention next validates the results in Example 1 at the protein level. The specific operation steps are as follows: put 2.0×10 5 LNCaP cell suspension per mL was seeded in 6-well plates according to 2 ml per well. The medium used was RPMI 1640 medium containing 10% activated carbon-treated fetal bovine serum (FBS); cells were cultured in a carbon dioxide incubator at 37 degrees Celsius. Cells were cultured for 48 hours after drug addition, 8 μL of DMSO was added to the solvent group, 4 μL of DHT with a final concentration of 5 nM was added to each well of the other groups, and DMSO, enzalutamide, and...
Embodiment 3
[0041] Example 3 ZJU-IMB-Z19 can inhibit the transcriptional activity of endogenous AR in LNCaP cells at the mRNA level.
[0042] The effect of ZJU-IMB-Z19 on AR transcriptional activity is mainly reflected in the effect of ZJU-IMB-Z19 on the expression of AR downstream target genes. PSA (KLK3) is an important downstream target gene of AR. In Example 2, the effect of ZJU-IMB-Z19 on PSA protein expression was detected. There may be two reasons for the decrease in PSA protein expression. On the one hand, it may be due to AR transcription. The activity is inhibited, resulting in the down-regulation of AR target gene expression, which may also be caused by the degradation of PSA protein on the other hand. In order to further confirm that the down-regulation of PSA is due to the inhibition of AR transcription, the present invention uses qRT-PCR method to detect the effect of ZJU-IMB-Z19 on PSA at the mRNA level.
[0043] The pretreatment method of this experiment is exactly the sa...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More