Hi everyone,
I would like to induce EMT in MCF7 cell line by TGFbeta1. From the literature, TGFbeta1 alone seems to induce a partial EMT even for one-week treatment. And this effect is concentration-dependent. What puzzles me most is that TGFbeta1 inhibits the mRNA expression of my gene of interest but up-regulates the expression level of protein. If this is really the case and if this is possible, what can be the mechanism?
Thanks a lot.
Xianqing
Hi Mao,
we tested TGFbeta effect in A549 cells. Below are two papers where we looked at effects on mRNA expression of Ah receptor and CYP1a1. The IC50 was only 5 pM TGFbeta1 in that case.
Good luck!
Chris
Effect of transforming growth factor-beta1 on expression of aryl hydrocarbon receptor and genes of Ah gene battery: clues for independent down-regulation in A549 cells.
Döhr O, Sinning R, Vogel C, Münzel P, Abel J.
Mol Pharmacol. 1997 May;51(5):703-10.
Transforming growth factor-beta 1 inhibits TCDD-induced cytochrome P450IA1 expression in human lung cancer A549 cells.
Vogel C, Döhr O, Abel J.
Arch Toxicol. 1994;68(5):303-7.
To Christoph,
Thanks for your answer but I cannot download your paper. Can you put a PDF file here?
To Eswar,
Thanks a lot for your answer. I just have another question: You treat your cells with 10ng/ml TGFbeta1, and you starve your cells or not? I saw that some people do that and the others not. Since you treat your cells during 3 days, I suppose that you treat them directly after your transfection, right?
Xianqing
MCF7 line lacks expression of the TGFB2 receptor to which TGFb binds. It is methylated (our unpublished data). Hence the effect that would come from binding of the ligand would be very minimal. If you are looking at EMT since these cells move as a cohesive sheet scratch test would be the best way to do the same. The traditional transwell assays do not work very well for MCF7 line.
please find the Mol Pharmacol paper attached. I have to find the other one.
here is the other article about TGFbeta on gene expression in A549 cells
Hi,
Please can you checked post transcriptional and transnational modification efficiency of your gene and protein of interest.
I checked the mRNA level, for example, by using 10ng/ml TGFbeta treatment, I got no difference even down-regulation in mRNA level of my gene of interest, while in protein level I can get a clear up-regulation. And is this what you mean?
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