Pharmacologic Effects of a Synthetic Macamide Analogue (ZYPC) on A549 and HeLa Cells

Abstract

Lepidium meyenii L., known as the Maca plant, is acclaimed by the scientific community for its various health benefits in traditional medicine, including prevention of gastritis, depression, and cancer, as well as increasing fertility and improving sexual performance. Macamides (N‐benzylamides) are secondary metabolites derived from Maca that have demonstrated neuroprotective, anti‐oxidative, anti‐inflammatory, endocannabinoid system modulatory, and mild anti‐diabetic properties. The present study investigates the pharmacologic effect of N‐(8Z‐heptadecen‐1‐yl)‐O‐(3‐pyridylmethyl) carbamate (ZYPC), a synthetic macamide analog, on A549 non‐small cell lung cancer and HeLa cervical cancer cells. Both cell lines were treated with different concentrations of ZYPC (1–100 μM). After 24 h, cell viability was measured using a 3‐(4, 5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTS) colorimetric assay. ZYPC was found to significantly reduce cell viability in a dose‐dependent manner. The half maximal inhibitory concentration was calculated, using dose‐response non‐linear regression, as 38.94 μM for A549 cells and 38.35 μM for HeLa cells. Subsequently, a caspase 3 colorimetric assay was executed to study ZYPC‐induced reduction of cell viability through activation of apoptosis. HeLa cells were treated with 0, 30, 50, and 100 μM ZYPC and cell lysates were collected after 5 h incubation. A reagent containing DEVD‐pNA, a substrate for caspase 3, was added to the lysates, and incubated for 2 h at 37°C. Unexpectedly, ZYPC was found to have no effect on caspase 3 activity. Then, the involvement of a pro‐survival mechanism was assessed through AKT and phospho‐AKT (p‐AKT) chemiluminescent western blotting. Results obtained from treatment with 0, 10, 30, and 50 μM ZYPC for 24 h were inconclusive for total AKT and p‐AKT levels in both cell lines. Preliminary western blots of poly (ADP‐ribose) polymerase (PARP), a DNA repair enzyme, and its inactive cleaved form indicated constant expression levels of both proteins in A549 cells. Interestingly, there was an apparent increase in cleaved PARP in HeLa cells, which may suggest late‐stage apoptosis. However, further studies are needed to better elucidate the mechanism of anti‐cancer activity exhibited by ZYPC.Support or Funding InformationMCPHS University Summer Undergraduate Research Fellowship Program