Scientific basis for the project

Cancer cells grow near the site where uncontrolled proliferation first began, resulting in the formation of the primary tumour mass; however, only about 10% of cancer deaths are caused by primary tumours. Instead, 90% of patients die because of cancer growths at sites away from the site of the primary tumour, i.e. those that form a metastasis. The first step in the formation of a metastasis is for cancer cells to undergo a phenotypic change. In the case of adenocarcinoma, the ovarian cancer studied in this project, the cancerous epithelial cells transition into a mesenchymal state which are capable of motility. This allows the cells to breach the basement membrane on which epithelial cells reside, breaking out of the primary tumour mass. It is therefore important to understand what factors may drive this phenotypic transition and motility.

Some voltage-gated sodium channels (VGSCs) are expressed in tumour cells and have been linked to cancer malignancy, with evidence of effects on tumour growth, invasion and metastasis (Mao et al., 2019). This project investigates whether VGSCs influence the motility of human ovarian cancer cells by observing the degree of migration occurring with and without certain VGSC antagonists.

Brief summary of each session

Before students enter their first lab, they attend an introductory lecture about ovarian cancer and the role of cell migration in ovarian cancer. The first lab introduces the students to the in-vitro scratch assay and allows them to try preliminary experiments. They create their first scratches, apply a DMSO negative control, a positive control and then image the initial scratches. In the second lab, students take repeat images of the scratches to assess the degree of cell migration. The remainder of this second lab is spent analysing the images using ImageJ and planning exactly which drugs and controls they will use in their experiment.

In the third lab, students start their real experiments, performing the in-vitro scratch assay with their chosen voltage-gated sodium channel blockers and appropriate controls, before taking the initial images. In the fourth and final lab, the students will take repeat images of the scratches to assess the degree of cell migration. They will then analyse the images and perform the appropriate statistical analysis.

The second and fourth labs have to be carefully timed to run the same day as the first and third labs, respectively. This is to enable long enough for the cells to have migrated but not long enough for all of the scratches to have completely closed.

Learning objectives

Following the successful completion of the project, you will be better able to:

1. Develop an appropriate hypothesis based on the current scientific literature
2. Design an experiment that effectively tests a hypothesis
3. Identify and use appropriate negative and positive controls
4. Evaluate the appropriateness of an experimental protocol and make modifications to a protocol based on preliminary results
5. Use aseptic technique, a pipettor and an inverted microscope
6. Calculate and perform the dilutions of drugs
7. Choose and perform appropriate imaging and data handling techniques
8. Evaluate results appropriately and put them into context

If you decide to use this practical, please let us know about it by filling in this short form – it’s not a requirement but we’d love to hear how our ideas are being used!