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Curcuma longa and Wild Type p53 Gene Induced
Apoptosis in MDA-MB-231 Breast Cancer Cell Line
Objective and Application
to Biotechnology
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Conclusion
Results and Interpretation
Eight million people die every year due to cancer.
This research will be a step forward in treating
cancer, saving those lives. This experiment will
focus on treating cancer using antioxidant,
specifically Curcuma longa, and p53 gene. This
comparative investigation will draw parallels and
contrast the effectiveness of antioxidant and p53
gene individually and both treatments combined.
Such treatment can be easily implemented since it
will not harm normal cells but will selectively kill
cancer cells.
If an antioxidant is added to the cancer cells, then
oxidative stress will be reduced. This will activate
the inserted p53 gene. Hence, cell death will be
initiated by the antioxidant and inserted p53.
Figure 2: Free radicals are damaging the cell.
Figure 1: Free radicals are produced through several
day-to-day activities.
Materials and Methods
Figure 3: Images of cells with plasmid DNA before and after
the experiment under a fluorescent microscope.
Figure 4: High levels of oxidative stress deactivate the p53 protein
and activate NF-kB protein which makes the cell cancerous.
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➢ Obtain p53 Plasmid
➢ Count Dead Cells
➢ Grow Single Colony
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➢ Obtain DNA
➢ Culture Cells
Figure 5: The graph shows the average percentage of cell death by all the experimental groups and the controls. The data shown is an average
of three trials. The error bars show the respective standard deviation of each variable. Untreated cells had 0% cell death; hence, no error bar is
shown. With that, p-values by T-tests are shown. * signifies p <= 0.05, ** signifies p <= 0.01, and *** signifies p <= 0.005.
➢ Transfect DNA
Abstract and Personal Information
Variables & Controls
Dependent Variable:
Dead Cells (%)
Experimental
Groups
Constant:
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Amount of PBS
Amount of Trypsin
Amount of media
Amount of Trypan blue
Amount of DNA
Etc.
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Controls
Eight million people die every year due to cancer. Hence, this experiment will focus on treating cancer using Curcuma longa
and p53 gene. This comparative investigation will draw parallels and contrast the effectiveness of the antioxidant and p53 gene
individually and both treatments combined. Furthermore, a wild type p53 plasmid was bought from Addgene. Then, a single
colony was isolated after overnight growth. This colony was grown for another 24 hours in LB Broth with Ampicillin antibiotic.
Then, miniprep kit from Qiagen was used to isolate plasmid DNA from the bacteria. It was transfected into MDA-MB-231
breast cancer cells using Lipofectamine 3000. Then, Curcuma longa (an antioxidant) was added after a day. After three days of
incubation, cells were counted using Trypan blue and a cell counter. In conclusion, Curcuma longa activated the inserted p53
protein and may have deactivated the NF-kB protein. Thus, apoptosis was induced in MDA-MB-231 breast cancer cell line.
Curcuma longa with p53 protein killed 56.67% cells on average. On the other hand, Curcuma longa and p53 gene individually
killed only 53.00% and 6.33% of cells respectively. The T-test confirms that most experimental groups were significantly
different from the controls.
Vraj Shroff, Niles West High School, Skokie, Illinois
Antioxidant initiates cell death in cancer cells
by altering transcription of proteins that induce
apoptosis and prevent cells from proliferating forever.
Cancer cells are very different from the normal cells
because of the high level of oxidative stress. This
inhibits the activities of p53 gene, hence apoptosis is
avoided. With that, NF-kB protein is overly expressed,
which allows cancer cells to proliferate infinitely.
However, antioxidants reduce oxidative stress in the
cells and reverse this situation. Curcuma longa
activates the inserted p53 gene and deactivates the
NF-kB protein. Thus, apoptosis is induced in
MDA-MB-231 breast cancer cell line by Curcuma
longa and p53 gene.
The results confirm the hypothesis of this
investigation. Curcuma longa and p53 protein killed
56.67% cells on average. On the other hand, Curcuma
longa and p53 gene individually killed only 53.00%
and 6.33% of cells respectively. The T-test confirms
that two experimental groups are significantly
different from their controls. The p-value of p53 DNA
with Curcuma longa to Control DNA with DMSO is
0.002. While p-value for Curcuma longa to DMSO is
0.023. Lastly, the p-value for p53 DNA to Control DNA
is 0.321. Hence, two of them are well under 5%
similarity.
The data also suggests that experimental groups
may be different from each other as well. The p-value
for Curcuma longa to p53 DNA is 0.004, and the
p-value for p53 DNA with Curcuma longa to p53 DNA
is 0.005. However, the p-value for p53 DNA with
Curcuma longa to Curcuma longa is 0.461, meaning
more trials are needed to find a statistical difference
between those two experimental groups.
Acknowledgements
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I am very grateful to Niles West High School for providing
me an opportunity to conduct my independent
experiment. I want thank District 219 Township for
providing lab equipment and grants to carry out this
research. I acknowledge the efforts of science faculty,
namely: Ms. Swiercz, Mr. Ruber, Ms. Gleicher, Mr. Kim,
and Ms. Villa. With all the teachers, I want to thank Ms.
Arcus for providing all the materials required for this
experiment. Lastly, I am indebted to Ms. Schwarz, my
mentor, for providing help around the clock. This
experiment may not have been possible without her.