Team:Tongji Shanghai/proof
proof
Part 1. in vitro cytotoxicity test
- The results shows that AuNRs do not affect the metabolic activities of the cells up to 100μl/mg..And they are stably dispersed in serum-containing medium without aggregation. Therefore, we proved that the AuNRs used in this study do not cause any acute cytotoxic effect or aggregation during ciculaiton in in vivo applications.
part 2: fluorescent proving
- Our part require hTERT promoter to reach specifically targeting, so we have designed experiment to prove that the part works as we expected..
part 3: western blot
- Now that we have proved that the hTERT promoter is working, we need to further illustrate that hsp promoter will work. We use p53 as downstream gene instead of GFP, and run western blot to check if the P53 protein will express higher when heated. We use 293T cells for positive reference. Most tumor cells don’t have high expression of p53 protein in general because the gene is inhibited or mutated. HCC1937 is no exception. However, 293T cells have higher expression level of p53 naturally, which makes the result easier to be detected.
- From the result, we can see that p53 is higher expressed inside cell when heat shocked, both in HCC1937 and 293T. In comparison, both cells remaining untreated has lower expressed p53, but still detectable. The hsp-p53 system is proved working in tumor cells.
Part 4. Mice experiment
- As time is limited, the vivo experiment is still in the progress. But we are glad to have see some fantastic changes.
- The six-day tumor volume shows that after infected hsp-p53 and hTERT-p53, the tumors are more likely to be depressed. With AuNRs and NIR, the depression is enhanced. It is likely that the system we developed actually work in vivo experiment.
