BACKGROUND: Colorectal cancer incidence and mortality rates are rising and are still one of the most common cancers worldwide. Its risks are associated with the Western Lifestyle, important facets of this being increase physical inactivity and excess body weight. In sync with colorectal cancer’s increasing rates are obesity rates that are rising in both developed and developing countries. Obesity has been linked to several causes of death that includes cardiovascular disease and certain cancers. Studies have shown the benefits of exercise overtly in alleviating cardiovascular diseases but now have increasing evidence of its benefits in helping with cancer. Some studies have examined the effects of physical activity in colon cancer and have found a link between increase minutes of exercise weekly can slow cancer cell proliferation in colon crypts cells. Myokines are a newly discovered class of proteins that are produced by muscles and have multiple functions. One of those function is its anti-neoplastic activity against certain cancer cell types. Certain myokines evoke an anti-neoplastic effect on cancer, but there has not been an overwhelming agreement within the scientific community.
OBJECTIVE: To investigate the anti-proliferative effects of exercise myokines produced by C2C12 cells in colorectal cancer cell lines: HCT116 and HT29, by looking at changes in protein expression through western blot analysis and protein arrays.
METHODS: C2C12 cells were grown in cell culture plates and differentiated into myotubules. Myotubules were subjected to Electronic Pulse Stimulation using an apparatus called C-Pace by Ion Optix to generate myokines. Treat HCT116 and HT29 cell lines with exercise myokines and a control treatment and ultimately harvest their protein lysate for western blot analysis and protein array analysis.
RESULTS: HCT116 treated with exercise myokines with different dilutions (1:25, 1:50, 1:100) a demonstrated significantly decreased proliferation (p<0.0001). HT29 treated with exercise myokines (1:25 dilution) also demonstrated a significant decrease in proliferation (p<0.0001). Western blot analysis of HCT116 treated with exercise myokines showed significantly lower expression in pRb (p<0.05), PCNA (p<0.05), and cyclin D1 (p<0.05). Western blot analysis of HT29 treated with exercise myokines revealed significant expression downregulation in exercise myokine treatment in PCNA (p<0.05), p42/p44 (p<0.05), pRb (p<0.0001) and Cyclin D1 (p<0.05). Human Phospho-Kinase Array demonstrated downregulation of the following proteins in HT29 cells treated with exercise myokines: GSK-3α/β, STAT3, EGFR, p53, PDGF Rβ, Src, PRAS40, WNK1, and JNK 1/2/3. Lastly, HT29 treated with exercise myokines showed downregulation of the p-EGFR and pHGFR as a result of the Human Phospho-RKT Array.
CONCLUSION: Our data demonstrate that exercise myokines produced by Electric Pulse Stimulation of C2C12 cells have an anti-proliferative effect on colorectal cancer cell lines HCT116 and HT29. Further studies should be pursued to identifying and linking anti-proliferative properties to a specific myokine protein and confirming a pathway that this specific myokine exerts its effects.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/36594 |
| Date | 13 June 2019 |
| Creators | Lopez, Timothy Magno |
| Contributors | Chowdhury, Sanjib |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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