Investigation of the in vitro anticancer potential of Eryngium wanaturii Woronow plant extract

Turkey, located in the Mediterranean basin, one of the important genetic centers of the Eryngium L. genus, possesses a rich diversity of Eryngium, including endemic species. One of these species, Eryngium wanaturii Woronow, is an endemic taxon unique to our country and its phytochemical content and biological activities have not been systematically studied. This study aims to conduct a phenolic profile scan of methanol and aqueous extracts obtained from the above-ground parts of the endemic species E. wanaturii and to investigate the effects of these two extracts on the pancreatic cancer cell line. Methanol and aqueous extracts were prepared from the aboveground parts of the plant using an ultrasonic bath. The Folin-Ciocalteu technique was used to determine the total phenolic content of the extracts, and the results were expressed as gallic acid equivalents. The aluminum chloride technique was used to determine the total flavonoid content, and the results were expressed as quercetin equivalents. To determine the total flavonoid content, the aluminum chloride technique was used, and the results were expressed as quercetin equivalents. Additionally, the phenolic compounds of both extracts were screened by LC/MS-QTOF analysis. The total phenol content was 32,6495±0,0348 mg GAE/g in the MeOH extract and 22,9617±0,0691 mg GAE/g in the water extract, while the flavonoid content was 24.7762 ± 0.0208 mg GAE/g in the methanol (MeOH) extract was 24.7762 ± 0.0208 mg KE/g and in the water extract was 11.2553 ± 0.0026 mg KE/g. According to LC-MS Q-TOF analysis results, chlorogenic acid was found in the plant's methanol extract at 2.98±0.20 µg/mg, and a low amount (<0.5 µg/mg) of caffeic acid was detected. The anticancer effects of the plant's methanol (MeOH) and water extracts on MIA PaCa-2 pancreatic cancer cells were investigated in vitro. The cells were cultured in DMEM medium and evaluated using different methods. MTT analysis showed that both extracts exhibited dose-dependent cytotoxicity. The MeOH extract produced a significant effect at 150 µg/mL and above, while the water extract produced a significant effect at 500 µg/mL and above. The IC50 values were found to be 580.8 ± 18.5 µg/mL (MeOH) and 578.6 ± 11.3 µg/mL (water), respectively . The scratch-wound healing analysis showed that both extracts significantly inhibited cell migration and metastasis potential. qRT-PCR and Western blot results revealed that the MeOH and water extracts increased Bax expression (by 24.61% and 9.02%, respectively) while decreasing Bcl-2 expression (by 12.83% and 6.89%, respectively). These results were also confirmed at the protein level. Immunofluorescence staining and microscopy revealed apoptosis-specific changes (cell shrinkage, microtubule loss, increased fluorescence intensity). In conclusion, E. wanaturi above-ground methanol and aqueous extracts exhibit anticancer effects in pancreatic cancer cells through cytotoxicity, migration inhibition, and apoptosis induction, with the methanol extract showing stronger biological activity compared to the water extract.