The hippo pathway is a network of proteins that control the size of tissues and organs in all organisms. Its role is to balance cell division and programmed cell death (apoptosis) It does this by controlling the number of cells in an organ. This allows organs to grow to the correct size. There seems to be a correlation between hippo and cancer. Scientists studying seem to believe that if we fully understand this gene we might be able to further our understanding of cancer.
Many of the proteins that make up the Hippo pathway are called tumour suppressors. The tumour suppressors work by preventing cancerous cells from growing. A mutation in a tumour suppressors’ gene allows cancerous cells to grow. A mutation in the hippo pathway can cause a variety of phenotypes to occur, such as: overgrown tissue, folded skin, tumours, bulges, and wrinkles.
Our research project was focused on finding out the sequence of the Hippo gene in a mutated fly, and discovering mutations in the DNA that would then allow scientists to further understand the hippo mutation, with the aim to help with cancer research. Our project involved many new and exciting procedures as well as a range of ultra modern technology that was very different to the equipment we had used before. We learnt how to extract DNA, purify it, replicate it and read its sequence. Our aim was to uncover why the hippo gene does mutate, and what specific part of its sequence has mutated.
Today on the 14th of May we broke down some larvae. Our objective was to extract DNA from the larvae of a fruit fly. The process was very slow. We isolated the DNA from the rest of the specimen by crushing it, then we used enzymes to break it down. We then centrifuged it to separate the DNA from the rest of the non-usable material using a membrane. We learnt many things while we were doing this. We learnt that DNA is made out of 3 parts, first being the nitrogen base (ATCG), second was the phosphate which is made from phosphorus and oxygen, and lastly the sugar which was Deoxyribose. We learnt how to use the centrifuge, the incubator, the micropipette and the vortex. We learnt that DNA was negatively charged because of the phosphate and oxygen.
Today on the 15th of May we tested if the DNA was there and then purified the samples. We discovered how to use a PCR (Polymerase Chain Reaction) machine which was used to duplicate the DNA. Before we used this machine we had to prepare the Purified Genomic DNA (gDNA). To make gDNA we had to mix primers, nucleotides, polymerase, buffer and the original DNA. When it heated up to 95 degrees, the DNA split apart into two strands. At 52 degrees, the primers bind to specific parts of the DNA we wanted, and the rest of the nucleotides attach to their complementary base. At 72 degree the polymerase made a back bone that stuck the nucleotide together. This was the cycle the PCR machine used to replicate the DNA samples.
Today on the 16th of May we did a series of workshop rotations. One of these included looking at the fly genetics and inheritance while also looking at the phenotypes in the flies using a microscope. We also looked into bioinformatics where we learnt about amino acids and how they are different, for example hydrophobic amino acids are scared of water but hydrophilic means that they are attracted to water. We were also able to examine our control flies and mutated flies under an SEM (Scanning Electron Microscope) and capture images of our flies at extreme magnification.
On the 17th of May, we were lucky enough to experience a private tour of the Peter MacCallum Cancer Research Centre. We visited a few different regions of the building, with the first being a specific lab dedicated to experiments and testing on flies. In this lab we got the chance to look at the tumours in the fly larvae through a green light setting on a microscope. This allowed us to easily see the tumours as they had been mutated so they were visible under a green light. In the next section we visited we were able to look under another microscope that in which we were able to compare a normal tissue cell to that of a mutated cell. Next we viewed another scientist in action, growing cancerous cells in a controlled environment. Lastly on this great tour we ventured into a room where the studies where being done on zebra fish instead of fruit flies. Thus showing us how the diversity in science projects allows bigger things to be uncovered.