Anti-proliferative effect of inactivated clostridium sporogenes and conditioned media on melanoma cells

• Main Author: Ng, Josie Wan Ting
• Other Authors: Teoh Swee Hin
• Format: Final Year Project
• Language: English
• Published: 2014
• Subjects: DRNTU::Engineering; DRNTU::Science
• Online Access: http://hdl.handle.net/10356/60802
• Institution: Nanyang Technological University

The rising incidence and high mortality rates of melanoma, together with partial response to the current methods of cancer therapy create a pressing need to look into alternative forms of cancer therapy. Although past studies have involved the use of bacteria and its components in the treatment of cancer, limitations exist. With the use of obligatory anaerobic bacteria to treat tumours, there is often extensive oncolysis resulting in death of the host and associated levels of toxicity. Therefore, elimination of toxicity associated with live bacteria might be necessary to provide a safer and more effective treatment. In this study, the anti-proliferative effect of inactivated Clostridium sporogenes and conditioned media on melanoma cells was investigated in vitro. 24 h, 48 h and 72 h after the addition of the C. sporogenes and conditioned media, cell proliferation assays were conducted to quantify their anti-proliferative effect on the cancer cells, which were grown in 96-well plates. Fluorescent images of the cells were obtained with the use of LIVE/DEAD® Cytotoxicity Assay and quantified. It was established that both inactivated C. sporogenes and its conditioned media have a tumour-killing effect on the melanoma cells. Generally, the addition of higher amounts of bacteria and prolonged incubation times led to increased cell death and lower cell viability. The addition of the conditioned media to the cancer cells resulted in much less viable cells than addition of bacterial growth media mixed with cancer cell growth media, therefore the anti-tumour effect was attributed to enzymes present in the conditioned media and not the lack of nutrients.