The Effects of Annona Muricata on The mTOR Pathway of Cervical Cancer Cells

Abstract

Annona muricata, also referred to as the soursop plant, has been gaining prevalence in the medical world due to its anticancer properties. Studied in several different cancer cell lines, the mechanism to how this plant stops the proliferation of rapidly diving cancer cells is still unknown. There have been reported involvements of Annona muricata in various cancerous proteins and pathways such as EGFR, MAPK, AKT, PKB, Cyclin D1 and NF‐kB, but there still remains some question to its activity in other target proteins. The purpose of this study was to demonstrate the activation or deactivation of the mammalian target of rapamycin (mTOR) complex in the HeLa line of cervical cancer cells in the presence of Annona muricata. The mTOR activity was monitored using Western Blot analysis of the activator AKT, and the direct downstream substrate, p70S6K1, to conclude if the mTOR was active and phosphorylating in the cell or if it was inhibited in the presence of Annona muricata. AKT was used to test that each cell culture had mTOR present to be monitored. In order to compare the mTOR activity of Annona muricata to a control group, cells were infused with rapamycin. Rapamycin, a macrolide antibiotic, is known to inhibit the phosphorylation of mTOR, leading to cell death. If the Western Blot analysis of the rapamycin incubated HeLa cells matched the Western Blot analysis of the Annona muricata incubated HeLa cells, it could be concluded that Annona muricata possibly inhibits mTOR, leading to its anticancer properties.In order to isolate the anticancer active ingredient of Annona muricata, there needed to be an extract of this plant to allow for chemical separation and testing. An Annona muricata ethyl acetate (AMEA) extract was used as the total extract in this the project. The AMEA extract was exposed to a thin layer chromatography (TLC) plate and observed to separate into five different fractions. Each fraction was then extracted from the silica gel, purified, and tested using a MTS assay of cell viability to gauge its anticancer properties. Through the cell viability analysis conducted, it was demonstrated that the fraction 2 (F2) of the separated total extract contained the anticancer activity within Annona muricata. The F2 was then exposed to the HeLa cell line, as well as the rapamycin being used as a control, for a time point of 3 hours. Each group of cells was then tested using Western Blot analysis to monitor the activation or deactivation of mTOR. Through this analysis, it was concluded that Annona muricata did not inhibit the mTOR complex in HeLa cells, although it still exhibited some sort of inhibition on the complex. Further studies can be conducted to monitor further downstream responses of mTOR in the presence of Annona muricata, such as autophagy, to conclude the involvement of Annona muricata in the inhibition of mTOR, and the anticancer effect.Support or Funding InformationMCPHS UniversityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.