title: Allyl-isatin suppresses cell viability, induces cell cycle arrest, and promotes cell apoptosis in hepatocellular carcinoma HepG2 cells.
abstract: The anticancer effect of the newly synthesized isatin derivative, N-allyl-isatin (Allyl-I), was evaluated in vitro with human hepatocellular carcinoma HepG2 cells. Cell viability was detected by cell counting kit-8 (CCK8) assay. Acridine orange (AO)/ethidium bromide (EB) double staining was used to observe the cell morphology. Flow cytometry was used to assess the effects of Allyl-I on the cell cycle, apoptosis rate, and mitochondrial membrane potential (MMP). Western blot analysis was performed to detect the influence of Ally1-I on the expression of cytochrome c (cyt c), Bax, Bcl-2, and cleaved caspase-3. Allyl-I significantly inhibited HepG2 cell viability in a time- and dose-dependent manner. Allyl-I can induce cell cycle arrest in HepG2 cells at the G2/M phase. Apoptotic nuclear morphological changes were observed after AO/EB double staining. Fluorescein isothiocyanate-conjugated Annexin V (Annexin V-FITC) and propidium iodide (PI) double staining showed that the apoptotic rates significantly increased in the presence of Allyl-I. Rhodamine 123 staining indicated that Allyl-I can decrease the MMP. Allyl-I also altered the expression of mitochondrial apoptosis-related proteins. Protein levels of cyt c and cleaved caspase-3 were upregulated following Allyl-I treatment. By contrast, the Bcl-2/Bax ratio decreased. Results suggest that Allyl-I suppresses cell viability, induces cell cycle arrest, and promotes cell apoptosis in HepG2 cells. Furthermore, the induction of apoptosis might be correlated with the mitochondrial pathway.
authors.affiliation: Department of Cell Biology, Binzhou Medical University, Yantai 264003, Shandong Province, China.
authors.forename: Weihua
authors.initials: W
authors.lastname: Bian
authors.affiliation: Department of physical, Binzhou Medical University, Yantai 264003, Shandong Province, China.
authors.forename: Yukuan
authors.initials: Y
authors.lastname: An
authors.affiliation: Department of Blood Transfusion, Affiliated Hospital of Binzhou Medical University, Binzhou 256603, Shandong Province, China.
authors.forename: Huiqing
authors.initials: H
authors.lastname: Qu
authors.affiliation: Department of Cell Biology, Binzhou Medical University, Yantai 264003, Shandong Province, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Cell Biology, Binzhou Medical University, Yantai 264003, Shandong Province, China.
authors.forename: Junhou
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Cell Biology, Binzhou Medical University, Yantai 264003, Shandong Province, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Xu
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D056945:Hep G2 Cells; D006801:Humans; D007510:Isatin / Q000494:pharmacology*; D008113:Liver Neoplasms / Q000188:drug therapy / Q000378:metabolism; D008996:Monoamine Oxidase Inhibitors
chemical_list: D000970:Antineoplastic Agents; D008996:Monoamine Oxidase Inhibitors; D007510:Isatin
keywords: allyl-isatin; cell apoptosis; cell cycle; flow cytometry; mitochondrial apoptosis pathway