Speaker | Transcript |
| Evan Becker | Hello, this is the |
| 2019 BJU cancer research team. |
| My name is Evan Becker and along with me today are Luke Brown, |
| Emily Swagger, Rachel Westfall and we are all under the direction of Dr. Stephen Figard at the Department of Biology and Bob Jones University. |
| And our paper, our presentation is on the molecular weight characterization of an almond component cytotoxic to gastrointestinal cancer cell lines. |
| All right, for the introduction. |
| 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. |
| Previous research from the BJU cancer lab has shown promise by demonstrating that almonds have a cytotoxic effect on LoVo colorectal cancer in vitro. |
| 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. |
| The same almond extract was shown to induce a dose response and the human gastric adenocarcinoma cancer cell line AGS. |
| 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. |
| Passing our almond extract through molecular weight cut off filters of 100,000 Daltons and 5000 Daltons respectively, |
| we were able to determine that the molecules present in the almonds inducing the set of toxic effect must be smaller than 5000 Daltons. |
| 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. |
| I think we're ready. |
| Rachel Westphal | 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. |
| 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. |
| WST cell proliferation assay, we utilize that to determine absorbance with a plate reader and use that to calculate percent viability. |
| 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. |
| Can go to the next slide. Yeah. |
| Luke Brown | 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. |
| 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. |
| 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, |
| 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, |
| 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. |
| 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. |
| 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 |
| what molecular weight, we'd be looking at to establish what compound or compounds are responsible for this effect. |
| 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. |
| Now you see that both the 100,000 molecular weight cut off filter and less than 5000 molecular weight cut off filter |
| 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. |
| 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. |
| cyanidin, delphinidin, malvidin and petunidin. |
| As you can see both the extract and all these flavonoids were actually more effective than the 5-FU in this treatment. |
| 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. |
| Emily | 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. |
| 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 |
| 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. |
