Cispubicin Case Study

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Discussion PART A DISCUSSION For Part A of the practical, we carried out a MTT assay on two drugs: Cisplatin and Doxorubicin, to see their effects on a Cisplatin sensitive cell line: A2780 and a Cisplatin resistant cell line: A270 Cp70. Three replicates were carried out for each drug versus each cell line at various concentrations in the MTT assay, where we used a 96 well-plate design. The absorbance was read and compiled in a table. I then took the averages of the replicates, and used that data to plot a graph of the absorbance values against the drug concentrations. Looking at the graph of Cisplatin versus A2780 and CP70, one can see that Cisplatin resulted in lower average absorbance values in the Cisplatin sensitive cell line than in…show more content…
In other words: would doxorubicin be a good alternative chemotherapeutic in a cisplatin-resistant patient based on my results? To answer this question I compared the ratio of the IC50 value for the CP70 cell line to the A2780 cell line when exposed to Cisplatin or Doxorubicin. Cisplatin’s concentration had to be doubled to achieve the same decrease in cell viability, while Doxorubicin need about 1.7 times as much, to also decrease cell viability by the same amount (50%) in the resistant cells. This indicates that the A2780 cell line is also resistant to Doxorubicin treatment. However, in this practical we used much higher concentrations of Cisplatin than Doxorubicin. The highest concentration of Cisplatin used was: 100 µM compared to only 5 µM of Doxorubicin. A much smaller amount of Doxorubicin was required to reduce cell viability by 50% in the resistant cells compared to Cisplatin. Doxorubicin is supposed to have an IC50 of 1-10 µM and Cisplatin is known to have an IC50 of about 50 µM. Yet in this experiment I obtained higher IC50 values for Doxorubicin, but this was due to the smaller amount of the drug used. Overall though, it is evident from my results that the person is cross resistant to Doxorubicin (which is a normal occurrence). However, because a much smaller…show more content…
The picture in this report is a sample result obtained from the module co-ordinator. The endonuclease activity that occurs in apoptosis causes the formation of non-oligonucleosomal fragment lengths, which forms a ‘ladder pattern’ on the agarose gel. (PRAC INFO) Looking at the picture an example of a sample that underwent apoptosis is the final column. One can see individual bars of DNA that resemble the rungs of a ladder. This indicates that the cell in a highly regulated process, was broken down to individual parts without the expulsion of its cellular contents into one, large clump. In cells that suffered a traumatic death, called necrosis, the cell rapidly loses membrane permeability, and the chromatin and thus DNA is not aggregated and broken down into smaller fragments. The DNA is detected as Random Fragment lengths, which can be visualised as a DNA smear on agarose gel. An example of a sample that underwent necrosis is column 2, where a large grey smear can be seen. This indicates that the cell remains stayed together (no apoptotic bodies were formed), and the chromatin underwent mottled condensation, which are characteristics of necrosis. To conclude, it is important that anti-cancer chemotherapeutic drugs such as Cisplatin and Doxorubicin induce apoptosis in cancerous cells instead of necrosis. This prevents a major immune response, and pro-inflammatory

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