- 18 Mar, 2021
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Report Structure
Your report should answer the questions listed below regarding the analysis and interpretation of the above data, understanding of the experimental technique, and the broader scientific context in which these experimental data sit. You must number each of your answers clearly in your report. You must provide full answers in written paragraphs. Single word or single sentence answers will not be adequate. Your answers should be properly referenced to indicate the sources of the information you present and a reference list should be included at the end of your report (see details below).
Question 1
Two cancer cell lines (Panc-1 and BxPC3) have been used in this experiment. What are these cell lines? What are some of the key biological properties of these cells? What are the main differences between these two cell lines?
Question 2
What is an SRB assay? Discuss in detail the purpose of the assay, how it works, and how it would be carried out.
Question 3
What is the purpose of the blank well containing only Tris buffer in the spectrophotometry measurements? Why is it included in the spectrophotometry measurements? How is the absorbance data obtained from the blank wells (shown in table 1) then used in the analysis of the other absorbance results?
Question 4
Copy table 1 into your report.
Then prepare a second table containing normalised absorbance results (by subtracting the absorbance of the blank from every absorbance value in the table). Make sure tables are numbered and are given an explanatory title.
Question 5
Using the normalised absorbance values in your second table, calculate the absorbance mean and the standard deviation for each condition (Panc-1 NT, Panc-1 Ly, Panc-1 Rap, BxPC3 NT, BxPC3 Ly, and BxPC3 Rap). Create a third table of the mean absorbance and standard deviation values.
Create a column chart of the absorbance means in your third table and add errors bars of the standard deviation values. Give the figure a number and figure legend.
Question 6
Compare the following conditions using an unpaired t-test.
Panc-1 NT versus Panc-1 Ly
Panc-1 NT versus Panc-1 Rap
BxPC3 NT versus BxPC3 Ly
BxPC3 NT versus BxPC3 Rap
To do this, use the online calculator at http://graphpad.com/quickcalcs/ttest1/ and enter the mean absorbance, standard deviation (SD) and number of replicates (N=3 for this experiment as each condition was measured by spectrophotometry three times) for the two conditions (groups) that you want to compare.
Create a fourth table containing the p-values from these analyses.
Question 7
Why was a t-test the most appropriate statistical test to use for the analyses in question 6? What does the p-value actually indicate?
Question 8
Briefly explain the PI3K and mTOR signalling pathways and their relevance to pancreatic cancer.
Question 9
Using the data in your figure (see question 5) and the p-values from the t-test comparisons (see question 6), discuss the effect of Ly294002 and Rapamycin on the proliferation of Panc-1 cells and BxPC3 cells, commenting on any differences between the two cell lines. Discuss whether you think we should use these inhibitors for the treatment of pancreatic cancer in human patients.