I am currently studying mutations in the RRAS gene and plan to functionally characterize these mutations; specifically, determine their effects on certain cancer hallmarks. To do this, I will be cloning a mammalian expression vector with the RRAS mutants and transfect them to certain cell lines. I am looking for a way to see their effects on angiogenesis. It was suggested that I check the expression of Hif-1alpha as an angiogenesis marker.
Dear Cherry:
Is a bit of controversy in this subject. Lekas et al demonstrated that MVD, VEGF cytoplasmic immunoreactivity and HIF-1alpha immunoreaction were more prominent in in tumor cells than in normal cells. On the other hand Hvingel demonstrated that the ratio between stromal expression of CD34 is the best toe evaluate angiogenesis. I personally think that it will depend on the type of cells that you will use.
In case you use endhotelial cells, those should be a extremely purified. Just in case I am sending you a paper of mine to do so: https://link.springer.com/article/10.1007/BF02639378
Best of luck
Daniel
Thank you Dr. Gomez :) I will check the literature on CD34. I recently came across the human angiogenesis antibody array by R&D system. I'm not really sure if we can afford to use them; if not, we'll probably just use the most significant ones.
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