VEGF has been related to angiogenesis in cancer and it expression is reported to decrease after treatment with a potent anticancer agents.
In my experiment (gene expression) i am failing to get such response. Even my positive control that is a known drug with anticancer profile is also increasing the gene expression of VEGF.
Any research publications, suggestions and comments in the light of said experiment will be highly appreciable.
In my opinion, an anticancer agent should not necessarily reduce the expression of VEGF. Inhibition of angiogenesis can be achieved through the competition of the anticancer agent with the ligand and prevent its binding to the receptor.
Hello Gaurav.
It could be that cancer cells become more easily drug-resistant upon chemotherapy. And this VEGF upregulations are well doumented.
See below.
- Targeting Chemotherapy-induced VEGF Up-regulation by VEGF Antisense Oligonucleotides in HNSCC Cell Lines.
- Tumour endothelial cells acquire drug resistance in a tumour microenvironment.
Regards.
Hi,
Try to evaluate 6-10 potential reference gene, and choose the most stable reference gene/genes in your experiment. Try to use a Normfinder algorithm to validate reference gene in your experiment.
Good luck!
Hi,
Try to evaluate 6-10 potential reference gene, and choose the most stable reference gene/genes in your experiment. Try to use a Normfinder algorithm to validate reference gene in your experiment.
Good luck!
@Sang Ho Lee
Sir, Thanks for your kind reply!
I am getting very good inhibitory concentration (50) at very micro-molar range, also the molecule is eliciting very good response toward 3D culture and wound healing assay, but failing to show any favorable results toward gene expression studies. Could it be due to i am using sub IC50 dosage or else?
@Justyna Augustyniak ,
Thank for your kind reply!
Along with VEGF, i have also used five other genes in which moreover the situation is same.
Could you suggest me at what dose of drug should i perform my gene expression studies if my IC50 of that particular drug is 2.2 micromolars.
Should it be less than IC50 or more than that?
Hello Gaurav.
It should be better to determine IC50 in a monolayer culture due to cell death caused by tight cell interactions formed within the inside of the 3D aggregates. There is no reliable method to evaluate cell death in 3D aggregate culture. See the discussions on similar topics. https://www.researchgate.net/post/Measuring_cell_proliferation_in_3D_cell_cultures
https://www.researchgate.net/post/cell_confluency_in_3D_scaffold
Regards.
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