Download methodologies - AOS-HCI-2011-Research

Investigation of the Effects of St. John’s Wort on Cancer and Drosophila
HCI
Objective 1 : To find out which immune related proteins interact with St. John’s wort or
hypericin.
1) Feed 10 drosophila flies with St. John’s Wort for 48 hours. Feed another 10
drosophila flies with their normal food for 48 hours.
2) Grind both sets of 2 sets of drosophila separately in sterile saline.
3) Conduct SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) on
the 2 solutions. Compare the 2 gel samples.
4) Extract relevant parts of the gel and send for MALDI-TOF (matrix-assisted laser
desorption/ionization time of flight mass spectrometry) @ NUS
Objective 2 : To find out whether St. John’s wort affects cancer cell growth
1) Culture Drosophila cancer cell line.
2) Add different concentrations of St. John’s wort (at least 4 concentrations) to media
containing cells. Allow incubation.
3) Cell count to check rate of proliferation of Drosophila cancer cells
AOS
Objective 1: To find out whether St. John’s wort affects cancer cell growth
1) Culture colon cancer cell line.
2) Add different concentrations of St. John’s wort (at least 4 concentrations) to media
containing cells. Allow incubation.
3) Cell count to check rate of proliferation of colon cancer cells
Objective 2: To find out whether St. John’s wort interacts with norepinephrine, which is
found to be responsible for cancer cell growth.
4) Culture colon cancer cell line.
5) Add different concentrations of St. John’s wort (at least 4 concentrations) to media for
it to interact with norepinephrine (since norepinephrine is secreted out of the cell).
Allow incubation.
6) Add samples from the media into the ELISA, which has wells attached with specific
anti-bodies.
7) Allow anti-bodies to bind with the norepinephrine present in the samples. Rinse.
8) Add coloured/fluorescent anti-bodies to bind to the anti-body norepinephrine
complexes present in the wells.
9) Measure the fluorescence using the absorbance plate reader.
Notes
 AOS would most probably not be experimenting with hypericin.
 Liquid St. John’s wort which is water based would be used. (proteins in powdered
samples may have denatured during drying)