title: A novel adamantane thiadiazole derivative induces mitochondria-mediated apoptosis in lung carcinoma cell line.
abstract: The interaction of organic compounds with apoptosis regulatory proteins is an attractive field of research because of its relevance in the development of new chemotherapeutic agents for cancer treatment. Our group designed four new adamantane thiadiazole derivatives (ATDs). The four ATDs were theoretically tested for their binding affinities to a model of an apoptosis inhibitor protein using molecular modeling. ATD-4 which interacted with the highest binding affinity was synthesized and characterized. The in vitro cytotoxicity of ATD-4 against different cancer cell lines as well as normal cell line was determined and compared with 5-fluorouracil as a standard positive control. The lung carcinoma cell line that showed the highest cytotoxic activity due to ATD-4 treatment was chosen to further study if ATD-4 can perform its cytotoxic activity through the induction of apoptosis as expected from molecular modeling. Inducing apoptosis by ATD-4 in lung carcinoma cell line was assessed by various biochemical and morphological characteristics. Biochemically: The effect of ATD-4 on cell cycle and its ability to induce apoptosis were checked through flow cytometry. Caspase-3 activity was detected by a colorimetric method. Real time-polymerase chain reaction (q-PCR) was used to detect p53, caspase-3, bcl-2 and bax gene expression. Morphologically: Changes in cell surface morphology, granulation and average surface roughness were detected using atomic force microscopy (AFM). Cell shrinkage, increase in cytoplasmic organelles, changes in mitochondrial number and morphology, chromatin condensation, membrane blebbing and formation of apoptotic bodies were detected using transmission electron microscopy (TEM). The obtained results suggest that ATD-4 exerted its antitumor activity against A549 cells through the induction of the intrinsic (mitochondrial) apoptotic pathway.
authors.affiliation: Department of Biochemistry, El Sahel Teaching Hospital, Cairo, Egypt.
authors.forename: Ayat G
authors.initials: AG
authors.lastname: Ali
authors.affiliation: Department of Chemistry, Biochemistry Speciality, Faculty of Science, Cairo University, Giza, Egypt.
authors.forename: Magda F
authors.initials: MF
authors.lastname: Mohamed
authors.affiliation: Department of Chemistry, Organic Chemistry Speciality, Faculty of Science, Cairo University, Giza, Egypt.
authors.forename: Abdou O
authors.initials: AO
authors.lastname: Abdelhamid
authors.affiliation: Department of Chemistry, Biochemistry Speciality, Faculty of Science, Cairo University, Giza, Egypt. Electronic address: mervat_sayed@yahoo.com.
authors.forename: Mervat S
authors.initials: MS
authors.lastname: Mohamed
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000648:ultrastructure; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D062105:Molecular Docking Simulation; D000072417:Protein Domains; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D059807:S Phase Cell Cycle Checkpoints / Q000187:drug effects; D013830:Thiadiazoles / Q000138:chemical synthesis / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000037:antagonists & inhibitors*; D051028:bcl-2-Associated X Protein / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; C494826:BAX protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D013830:Thiadiazoles; D016159:Tumor Suppressor Protein p53; D051636:X-Linked Inhibitor of Apoptosis Protein; C496866:XIAP protein, human; D051028:bcl-2-Associated X Protein; C505407:CASP3 protein, human; D053148:Caspase 3; D000218:Adamantane; D005472:Fluorouracil
keywords: Adamantane thiadiazole derivatives; Apoptosis; Biochemical and morphological characteristics; Cytotoxicity; Molecular modeling