Alright everyone, I’ve been keeping you at the edges of your seats for long enough…maybe? Regardless, now that you more or less understand the theory behind my project, let’s actually get into the details. So I’ve been doing this project for how many weeks now? 6? Yet I haven’t really given you anything about the results or analysis, so that changes today.
Now based on my FACS results from 4 trials, which seem to be pretty precise, the best conditions are the 1.25 µM Dox, 1.25 µM Dox + Pac, and pretreatment of 1.25 µM Dox and Pac. Furthermore, it doesn’t look like the results from the pretreatment conditions are that significantly different from the Dox-only conditions. However, adding Pac to Dox seems to induced more apoptosis in the HepG2 cells than Dox alone. So since Pac targets liver cells and stops them at the G2/M checkpoint, apoptosis is induced in these cancer cells. Then Dox targets the rest of the cells, which are dividing. As a result, there are fewer HepG2 cells that can evade the effects of Dox since they go through apoptosis because of Pac. According to my Western Blot results, all the proteins are being expressed as they should, they all show an accurate trend (which will be shown and explained in more detail in the final product).
One thing I find weird however is that the last 3 conditions (2.5 µM Dox, 2.5 + Pac, and 2.5 pretreatment + Pac) don’t fit in with the trend. There’s significantly fewer apoptotic cells than the 1.25 conditions, and all the proteins are not being expressed at the levels they should be. So at first I thought maybe the concentration is too toxic, but I was wrong, because the regular concentration given to patients is over 20-30 µM. So now, I’m thinking of two theories: one being the drug decayed too fast and the cells had time to resist and repopulate (which seems a little less reasonable), and the other being that the cells switched to death because of elevated levels of ROS or oxidative stress. This is much different from apoptotic death. The latter seems much more reasonable to me, and I have to ask my professor once he gets back from India.
From now until the end of the 10 week period, I’ll just be conducting more trials for the sake of accuracy.