title: p53 dependent apoptosis and cell cycle delay induced by heteroleptic complexes in human cervical cancer cells.
abstract: We previously reported synthesis of novel arene ruthenium (Ru) complexes and evaluated their antitumor activity in murine lymphoma (DL) cells. In this present study we further investigated the mechanism of action of two ruthenium complexes [complex 1 (η6-arene)RuCl(2-pcdpm)] and complex 2 (η6-arene)RuCl(4-mtdpm)] in cervical cancer cell line (HeLa). Our studies demonstrate that anticancer property of these two complexes was due to induction of apoptosis through p53 mediated pathway as well as arrest of cells in G2/M phase of cell cycle. It is worth to note that the complexes did not cause any substantial cytotoxic effect on normal cells. Further in comprehensive studies, apoptosis inducing property of both complexes were established in accordance with array of morphological changes ranging from membrane blebbing to formation of apoptotic bodies and followed by DNA fragmentation assay. Furthermore, Flow cytometry by Annexin V/PI staining delineate that complex 1 and 2 have strident impact to induce apoptosis in HeLa cells. The complex 1 and 2 treated cells show increased level of intracellular ROS generation which was preceded by p53 activation. Apoptosis induced by 1 and 2 was preceded by mitochondrial aggregations which were monitored by mitotracker. In addition flow cytometry analysis showed that both complexes also effectively arrest cells at G2/M phase of cell cycle. Western blot, RT-PCR as well as Real Time analysis were used to further confirm that the complexes induced apoptosis in p53 dependent pathway. Thus, our promising results can contribute to the rational design of novel potential anticancer agents.
authors.affiliation: Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India. Electronic address: gunjansharmabhu10@gmail.com.
authors.forename: Gunjan
authors.initials: G
authors.lastname: Sharma
authors.affiliation: Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India. Electronic address: rananishant19nov@gmail.com.
authors.forename: Nishant Kumar
authors.initials: NK
authors.lastname: Rana
authors.affiliation: Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India. Electronic address: priyasinghbhu20@gmail.com.
authors.forename: Priya
authors.initials: P
authors.lastname: Singh
authors.affiliation: Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India. Electronic address: gauravcdubey91@gmail.com.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Dubey
authors.affiliation: Department of Chemistry, Institute of Science, Banaras Hindu University, Varanasi, 221005, India. Electronic address: dspbhu@bhu.ac.in.
authors.forename: Daya Shankar
authors.initials: DS
authors.lastname: Pandey
authors.affiliation: Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India. Electronic address: kochbiplob@gmail.com.
authors.forename: Biplob
authors.initials: B
authors.lastname: Koch
mesh_terms: D000072283:A549 Cells; D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D059447:Cell Cycle Checkpoints / Q000235:genetics / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000473:pathology / Q000648:ultrastructure; D053938:DNA Fragmentation; D005260:Female; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000473:pathology / Q000648:ultrastructure; D017382:Reactive Oxygen Species / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000502:physiology*; D002583:Uterine Cervical Neoplasms / Q000235:genetics*
chemical_list: D018797:Cell Cycle Proteins; D017382:Reactive Oxygen Species; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53
keywords: Anticancer; Antiproliferative activity; Apoptosis; Cell cycle; Cytotoxicity; Reactive oxygen species