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Speaker
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Transcript
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Evan Becker
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Hello, this is the
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2019 BJU cancer research team.
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My name is Evan Becker and along with me today are Luke Brown,
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Emily Swagger, Rachel Westfall and we are all under the direction of Dr. Stephen Figard at the Department of Biology and Bob Jones University.
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And our paper, our presentation is on the molecular weight characterization of an almond component cytotoxic to gastrointestinal cancer cell lines.
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All right, for the introduction.
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Colorectal cancer is currently of great concern in the medical community as it is the third leading cause of cancer deaths for both men and women nationwide.
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Previous research from the BJU cancer lab has shown promise by demonstrating that almonds have a cytotoxic effect on LoVo colorectal cancer in vitro.
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A pseudo digestion procedure for an almond extract was also used as a way to mimic how the extract would work in a physiological system.
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The same almond extract was shown to induce a dose response and the human gastric adenocarcinoma cancer cell line AGS.
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Also this extract causes no negative effects in the human colon epithelial cell line CCD, indicating that the cytotoxic effects of almonds do not affect normal healthy cells.
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Passing our almond extract through molecular weight cut off filters of 100,000 Daltons and 5000 Daltons respectively,
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we were able to determine that the molecules present in the almonds inducing the set of toxic effect must be smaller than 5000 Daltons.
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As a result, polyphenols were determined to be a possible cause of the cytotoxic effect and the polyphenol extract was conducted on the almonds with this treatment showing very similar cytotoxic effects on the cancer cell lines.
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I think we're ready.
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Rachel Westphal
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To the cell lines that we use included AGS, which is stomach cancer; LoVo, which is colon cancer; and CCD, which was our normal cell line.
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We used 5-fluorouracil as our positive control for cell death. And we used an in vitro pseudo digestion of the almonds to mimic physiological digest...physiological digestion.
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WST cell proliferation assay, we utilize that to determine absorbance with a plate reader and use that to calculate percent viability.
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And then JMP was used for statistical analysis, we used ANOVA analysis, the Tukey-Kramer HSD test and the Wilcox and non parametric comparison. p values less than 0.05 were considered statistically significant.
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Can go to the next slide. Yeah.
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Luke Brown
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All right. The first set of tests I'd like to introduce you to are tests regarding the AGS human cancer cell line and this is a human gastric adino carcinoma.
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Now JMP was an important tool for us because it helped us to, first of all, determine the standard deviation in a few pilot studies we conducted.
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This then allowed us to use the software to run a power analysis to determine our sample size. Now looking at some of the data we got here,
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first of all, you'll see among both Figure 1 and Figure 2 what it has in common is PBS phosphate buffered saline and 5-FU or 5-fluorouracil,
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which is well documented and established cancer treatment for cancer, such as gastric and colorectal cancers. Now I'd like to direct your attention to Figure 1 here.
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Previous studies in our lab had already established that it seems almond extract does have some sort of cytotoxic effect that's selective to these cancer lines.
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Well, we hope to establish in Figure 1 here is, first of all, the dosage effect. And second of all, we wanted to narrow down
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what molecular weight, we'd be looking at to establish what compound or compounds are responsible for this effect.
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As you see, moving up from 12% almond extract all the way to 100% almond extract, we do see a dosage response. And we were able to establish this using the Tukey-Kramer HSD test inside JMP to be able to establish that these groups are statistically significantly different.
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Now you see that both the 100,000 molecular weight cut off filter and less than 5000 molecular weight cut off filter
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are statistically the same as the 100% almond extract. This led us to believe that whatever compound or compounds is responsible for this effect is going to be relatively small at less than 5,000 Daltons.
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Given this, we moved on to Figure 2 and we looked at some research showing that flavonoids have been shown to have a similar effect in walnuts.
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cyanidin, delphinidin, malvidin and petunidin.
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As you can see both the extract and all these flavonoids were actually more effective than the 5-FU in this treatment.
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So given this information, I'm going to pass the next slide to Emily here to give you some more information about another set of data.
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Emily
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So to make sure that we weren't just looking at results particular to AGS, we also ran another cancer cell line called LoVo. LoVo is a colon cancer cell line.
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As you can see, we didn't do all of the extensive dosage treatments on this particular one, because we'd kind of shown that with AGS. We were just particularly looking at
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is this particular to a certain line? So for LoVo, you can see that we don't have as low a value for 5-FU.
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