癌症药理学:Provide recommendations in a multi-million dollar project

In this simulation, you will learn which tools are needed to screen thousands of chemical compounds in order to identify promising candidates for the treatment of different types of cancer. In particular, you will gain insight into the principles and limitations of widely used in vitro assays in the context of anti-cancer drug screenings. As a pharmacology expert, you will test the robustness of those commonly used techniques and help a screening facility to assess the risk for false negative results.

Prepare cells and drugs for chemosensitivity testing

Your main task in the Cancer Pharmacology simulation is to reproduce the screening facility’s chemosensitivity in vitro protocol to investigate the cause of false negative results. The protocol in question is a widely used cell survival assay using MTT reagent and performed with conventional cancer cell lines. The Project Leader suggests to perform the test with two well-known chemotherapy drugs: epirubicin and cyclophosphamide. In this part of the simulation, you will learn how to perform common steps of chemosensitivity testing and learn about basic pharmacology tools. Most importantly, you will count living cells of a cell culture solution and calculate how much of the culture you need for your experiment. You will also prepare stock solutions of both drugs with a given concentration to create a dilution series for the treatment of the cancer cells.

Perform the MTT assay

一旦准备好细胞并与药物一起孵育，就该将MTT用作细胞存活指标了。您将能够在分子水平上遵循MTT如何与线粒体酶特异性反应，从而导致可以用分光光度计测量的颜色比色变化。在此任务中，您将使用多通道移液器和96张井板，以执行全球许多药理学实验室中使用的测定。

确定EC50并解释您的结果

The result of the serial drug dilution in the MTT assay will provide the basis for creating a dose-response curve and for determining the half-maximal effective concentration (EC50) of both clinically used drugs. Which recommendations will you provide the Project Leader with about the usefulness of the commonly applied in vitro assays?