I am planning to find the Hub protein using protien-protein interaction through in-sillico apporach. For that which is the most suitable cell lines(tumor).
I agree with Ruhul. You can not induce EMT in MCF7 using TGF-beta. An excellent model includes MCF10A and HMLE (but they have very strict and complex culturing conditions). An amazing model I've been using for my studies is NMuMG cell lines. They respond very quickly to TGF-beta completely changing their morphology in as little as 4-5 days in 10ng/mL of TGF-beta. You start to see reduction of epithelial markers at the mRNA level in 24 h and 48 h at the protein level. HMLE in my opinion is a bit slower to see morphological changes (6-8 days). NMuMG culture media is really simple (DMEME + Insulin) and they are available on ATCC unlike HMLE. Good luck!
Hello,
It depends where (which tissue) you want to investigate the TGF-induced EMT effect. If it required to be tumor, I could suggest HepG2 in liver or MDA MB for breast tumor (this could be careful with maglinant characteristic of MDAMB in EMT), or TGF even works very well with HCT116 colorectal cancer cell line. See this link: http://www.e-crt.org/journal/view.php?number=2521. You can also test in normal cell line like Chang, Huh7 (hepatocyte) which also gave the clear EMT effect with TGF treatment. Or with keratinocyte HaCaT also works.
Hope it helps :) Good luck!
MDAMB231 is one of the most known metastasis cell lines of breast cancer. You can also try PC3, DU145 and LNCAP of prostrate cancer.
I guess cell lines with high metastatic potential can be used for the study of TGF-beta induced EMT. In that case, one of the highly metastatic potential cell line with extensive studies is MDA-MB-231. You can go through the following links for your reference.
http://www.sciencedirect.com/science/article/pii/S0167488911002151
http://www.nature.com/onc/journal/v21/n49/full/1205966a.html
I am very thank full for your kind suggestions
Mr. Manh Tin Ho and Mr.Iqbal Mahmud
I am planning to check in MCF-7 cell lines and replicate the same in in-vivo EAC model system would it be suitable for my study ?
TGFbeta may not induce EMT in MCF7 cell because of the repressed state of chromatin in EMT inducer gene's promoter (http://www.ncbi.nlm.nih.gov/pubmed/23827675). In fact, I could not induce EMT in MCF7 by TGFbeta. MDA-MB-231 is a cell line which has already been in mesenchymal state, so it will not be a good choice. Because you may not see the morphological difference upon TGFbeta treatment. You need some cell lines which are epithelial, express lot of E-cadherin. One choice can be MCF10A or HMLE cell line. You can look at the following paper (http://www.cell.com/cell/pdf/S0092-8674%2808%2900444-3.pdf)
I am very thank full for your kind suggestions
Mr. C.R. Naveen Kumar and Mr.Ruhul Amin
I agree with Ruhul. You can not induce EMT in MCF7 using TGF-beta. An excellent model includes MCF10A and HMLE (but they have very strict and complex culturing conditions). An amazing model I've been using for my studies is NMuMG cell lines. They respond very quickly to TGF-beta completely changing their morphology in as little as 4-5 days in 10ng/mL of TGF-beta. You start to see reduction of epithelial markers at the mRNA level in 24 h and 48 h at the protein level. HMLE in my opinion is a bit slower to see morphological changes (6-8 days). NMuMG culture media is really simple (DMEME + Insulin) and they are available on ATCC unlike HMLE. Good luck!
A549 pulmonary adenocarcinoma cells are a very tractable cell model for investigating TGF-beta-induced EMT. After 24-hour incubation with 5 ng mL-1 TGF-beta 1, there is a robust repression of E-cadherin/increase in fibronectin expression. Also very easy to see morphological changes via bright-field microscopy.
https://www.ncbi.nlm.nih.gov/pubmed/15084245
I recommend this paper. The best lines to see EMT with, as suggested, are NMuMG and MCF-10A cells for mammary epithelia. This paper has additional lines you can try for other systems. MCF-7 unfortunately has mutated TGFBRII receptors and will not respond to TGFb induction. There are supposedly some lines out there that do respond to TGF-beta- but not the one from ATCC.
- Badges
- Science topic