Vpliv izbranih prostih in v nanodelce vgrajenih antagonistov nikotinskih acetilholinskih receptorjev na celice raka pljuč

Joukhan, Ahmad (author), Drobne, Damjana (mentor), Sollner Dolenc, Marija (comentor)

The overexpression of nicotinic acetylcholine receptors (nAChRs), particularly ?7-nAChR, in lung cancer cells facilitates the binding of nicotine or other nAChR agonists such as nicotine metabolites or acetylcholine. The binding and activation of these receptors not only promote tumor growth but also enhance cancer resistance to chemotherapy. This study investigated the efficacy of various nAChR antagonists (APS7-2, APS12-2 and APS8-2) in attenuation of the nicotine effects on A549 lung cancer cells. APS7-2, APS8-2, and APS12-2 are synthetic analogs of the marine sponge toxin 3-alkylpyridinium salts (poly-APS). APS12-2 acts as a ?12ß1?? nAChR antagonist, while our electrophysiological experiments demonstrated that APS7-2 and APS8-2 function as ?7-nAChR antagonists. We also investigated the potential of APS12-3, another poly-APS analog, to attenuate nicotine effects on A549 cancer cells. APS8-2 did not exhibit cytotoxicity on A549 cells, whereas APS7-2, APS12-2, and APS12-3 demonstrated cytotoxicity. The effects of nicotine on A549 cells include heightened cell viability and proliferation, elevated levels of Ca2+, and diminished production of reactive oxygen species (ROS) induced by cisplatin. These effects were attenuated by APS8-2, APS12-2, and APS12-3, while APS7-2 was less effective. APS12-2-loaded gelatin nanoparticles (APS12-2-loaded GNPs) were prepared using the nanoprecipitation technique, and their cytotoxic effects were assessed on A549 cells and BEAS-2B non-tumorigenic lung cells. The cytotoxicity of APS12-2 on BEAS-2B cells was significantly reduced when APS12-2 was loaded into GNPs, whereas the cytotoxicity on A549 cells was only slightly decreased. The potential of APS12-2 and APS12-2-loaded GNPs was also investigated on nicotine-induced reduction in cisplatin efficacy. Nicotine reduced the cytotoxicity, formation of ROS, and lipid droplet formation induced by cisplatin in A549 cells. APS12-2 and APS12-2-loaded GNPs attenuated the nicotine-induced reduction in the efficacy of cisplatin. Our findings indicate that both APS12-2 and APS12-2-loaded GNPs hold promise as supportive agents in chemotherapeutic cisplatin for lung cancer.

• Language: English
• Keywords: marine toxins, nicotinic acetylcholine receptors, nAChR antagonists, 3-alkylpyridinium salts, chemotherapeutic agents
• Work type: Doctoral dissertation
• Organization: FFA - Faculty of Pharmacy
• Year: 2024
• PID: 20.500.12556/RUL-161633
• Publication date in RUL: 13.09.2024

Secondary language

• Language: Slovenian
• Title: The effects of selected free and nanoparticles embedded antagonists of nicotinic acetylcholine receptors on lung cancer cells

Abstract

Povečana ekspresija nikotinskih acetilholinskih receptorjev (nAChR), še posebej α7-nAChR, v celicah pljučnega raka povečuje vezavo nikotina ali drugih agonistov nAChR, kot so metaboliti nikotina ali acetilholina. Vezava na te receptorje in njihova aktivacija spodbujata rast tumorja ter povečujeta odpornost na kemoterapijo. V tej raziskavi smo preučili potencial različnih antagonistov nAChR (APS7-2, APS8-2 in APS12-2) pri preprečevanju učinkov, ki jih povzroča nikotin na celice pljučnega raka A549. APS7-2, APS8-2 in APS12-2 so sintetični analogi toksinov poli-3-alkilpiridinijevih soli (poly-APS), ki izhajajo iz morske spužve. APS12-2 deluje kot antagonist α12β1γδ nAChR, medtem ko so naši elektrofiziološki poskusi pokazali, da APS7-2 in APS8-2 delujeta kot antagonista α7-nAChR. Prav tako smo preučili potencial APS12-3, drugega analoga poly-APS, pri zaviranju učinkov nikotina na celice A549. APS8-2 (0.5-100 µg/mL) ni pokazal citotoksičnih učinkov na celice A549, medtem ko sta APS7-2 (1 µg/mL), APS12-2 (0.5 µg/mL) in APS12-3 (1 µg/mL) pokazala citotoksičnost. Z nikotinom povzročeni učinki na celice A549 vključujejo povečanje živosti in proliferacije celic, dvig ravni Ca2+ ter zmanjšano proizvodnjo reaktivnih kisikovih zvrsti (ROS), ki jih povzroči cisplatin. Te nikotinske učinke smo zmanjšali z APS8-2, APS12-2 in APS12-3, medtem ko je bil APS7-2 manj učinkovit. Nanodelci z APS12-2 (APS12-2-GNPs) so bili pripravljeni s tehniko nanoprecipitacije, njihovi citotoksični učinki pa so bili preizkušeni na celicah A549 in netumorskih celic pljuč BEAS-2B. Citotoksičnost APS12-2 na celice BEAS-2B je bila znatno zmanjšana, ko so bile izpostavljene APS12-2-GNPs, medtem ko je bila citotoksičnost na celice A549 le rahlo zmanjšana. Prav tako smo preučili potencial APS12-2 in APS12-2-loaded GNPs v zvezi z nikotinsko-induciranim zmanjšanjem učinkovitosti kemoterapevtika cisplatina. Nikotin je zmanjšal citotoksičnost, tvorbo ROS in tvorbo lipidnih kapljic, ki jih povzroči cisplatin v celicah A549. APS12-2 in APS12-2-loaded GNPs sta zmanjšala nikotinsko inducirano zmanjšanje učinkovitosti cisplatina. Naša raziskava kaže, da imata tako APS12-2 kot tudi APS12-2-loaded GNPs potencial kot podporno sredstvo pri kemoterapiji s cisplatinom pri pljučnem raku.

• Keywords: morski toksini, nikotinski acetilholinski receptorji, antagonisti nAChR, 3-alkilpiridinijeve soli, kemoterapevtiki