A novel bispidinone analog induces S‑phase cell cycle arrest and apoptosis in HeLa human cervical carcinoma cells

Yi,Xu; Zhang,Xin; Jeong,Hyunjin; Shin,Yu Mi; Park,Dong Ho; You,Song; Kim,Dong-Kyoo

doi:10.3892/or.2015.3722

A novel bispidinone analog induces S‑phase cell cycle arrest and apoptosis in HeLa human cervical carcinoma cells

Authors:
- Xu Yi
- Xin Zhang
- Hyunjin Jeong
- Yu Mi Shin
- Dong Ho Park
- Song You
- Dong-Kyoo Kim
View Affiliations

Affiliations: Department of Chemistry and Institute of Basic Sciences, Inje University, Gimhae 621-749, Republic of Korea, School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, P.R. China
Published online on: January 15, 2015 https://doi.org/10.3892/or.2015.3722
Pages: 1526-1532

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

2,4,6,8-(3)-Tetranitrophenyl-3,7-diazabicyclo[3.3.1]nonan-9-one (B16), a bispidinone analog, was synthesized to investigate its effects on cell viability, the cell cycle, and apoptotic pathways in HeLa human cervical cancer cells. B16 decreased the percentage of viable cells in WST-8 assays, and morphological changes associated with apoptotic cell death were observed, including cell shrinkage and disruption. Annexin V-FITC/PI dual staining assays showed that B16 significantly increased the early apoptosis of HeLa cells after 24 h of treatment. Moreover, DNA content analysis and [3H]-thymidine incorporation assays showed that B16 induced S-phase cell cycle arrest and inhibited DNA replication after 24 h of treatment. Following treatment with 25 µM of B16, an increase in reactive oxygen species and a decrease in mitochondrial membrane potential were observed by flow cytometry. In addition, the expression levels of caspase cascade and Bcl-2 family proteins determined by western blotting suggested that the induction of apoptosis by B16 was associated with a caspase- and mitochondrial-dependent pathway in HeLa cells. In conclusion, B16 induced early apoptosis and S-phase cell cycle arrest in HeLa cells via a caspase- and mitochondrial‑dependent pathway.

View Figures

View References

1	Bray F, Jemal A, Grey N, Ferlay J and Forman D: Global cancer transitions according to the Human Development Index (2008–2030): a population-based study. Lancet Oncol. 13:790–801. 2012. View Article : Google Scholar : PubMed/NCBI
2	Smith HO, Tiffany MF, Qualls CR and Key CR: The rising incidence of adenocarcinoma relative to squamous cell carcinoma of the uterine cervix in the United States - a 24-year population-based study. Gynecol Oncol. 78:97–105. 2000. View Article : Google Scholar : PubMed/NCBI
3	Rein DT and Kurbacher CM: The role of chemotherapy in invasive cancer of the cervix uteri: current standards and future prospects. Anticancer Drugs. 12:787–795. 2001. View Article : Google Scholar : PubMed/NCBI
4	Gascoigne KE and Taylor SS: Cancer cells display profound intra-and interline variation following prolonged exposure to antimitotic drugs. Cancer Cell. 14:111–122. 2008. View Article : Google Scholar : PubMed/NCBI
5	Jemal A, Center MM, DeSantis C and Ward EM: Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol Biomarkers Preg. 19:1893–1907. 2010. View Article : Google Scholar
6	Sherwood SW, Sheridan JP and Schimke RT: Induction of apoptosis by the anti-tubulin drug colcemid: relationship of mitotic checkpoint control to the induction of apoptosis in HeLa S3 cells. Exp Cell Res. 215:373–379. 1994. View Article : Google Scholar : PubMed/NCBI
7	Evan GI and Vousden KH: Proliferation, cell cycle and apoptosis in cancer. Nature. 411:342–348. 2001. View Article : Google Scholar : PubMed/NCBI
8	Pucci B, Kasten M and Giordano A: Cell cycle and apoptosis. Neoplasia. 2:2912000. View Article : Google Scholar : PubMed/NCBI
9	Xiao J, Huang G, Zhu C, Ren D and Zhang S: Morphological study on apoptosis HeLa cells induced by soyasaponins. Toxicol In Vitro. 21:820–826. 2007. View Article : Google Scholar : PubMed/NCBI
10	Dewson G and Kluck RM: Mechanisms by which Bak and Bax permeabilise mitochondria during apoptosis. J Cell Sci. 122:2801–2808. 2009. View Article : Google Scholar : PubMed/NCBI
11	Zamzami N and Kroemer G: The mitochondrion in apoptosis: how Pandora’s box opens. Nat Rev Mol Cell Biol. 2:67–71. 2001. View Article : Google Scholar : PubMed/NCBI
12	Simizu S, Takada M, Umezawa K and Imoto M: Requirement of caspase-3(-like) protease-mediated hydrogen peroxide production for apoptosis induced by various anticancer drugs. J Biol Chem. 273:26900–26907. 1998. View Article : Google Scholar : PubMed/NCBI
13	Liu MJ, Wang Z, Li HX, Wu RC, Liu YZ and Wu QY: Mitochondrial dysfunction as an early event in the process of apoptosis induced by woodfordin I in human leukemia K562 cells. Toxicol Appl Pharmacol. 194:141–155. 2004. View Article : Google Scholar : PubMed/NCBI
14	Kluck RM, Bossy-Wetzel E, Green DR and Newmeyer DD: The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science. 275:1132–1136. 1997. View Article : Google Scholar : PubMed/NCBI
15	Putcha GV, Harris CA, Moulder KL, Easton RM, Thompson CB and Johnson EM: Intrinsic and extrinsic pathway signaling during neuronal apoptosis lessons from the analysis of mutant mice. J Cell Biol. 157:441–453. 2002. View Article : Google Scholar : PubMed/NCBI
16	Day TW, Huang S and Safa AR: c-FLIP knockdown induces ligand-independent DR5-, FADD-, caspase-8-, and caspase-9-dependent apoptosis in breast cancer cells. Biochem Pharmacol. 76:1694–1704. 2008. View Article : Google Scholar : PubMed/NCBI
17	Graña X and Reddy EP: Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases (CDKs), growth suppressor genes and cyclin-dependent kinase inhibitors (CKIs). Oncogene. 11:211–219. 1995.PubMed/NCBI
18	Hall M and Peters G: Genetic alterations of cyclins, cyclin-dependent kinases, and Cdk inhibitors in human cancer. Adv Cancer Res. 68:67–108. 1996. View Article : Google Scholar : PubMed/NCBI
19	Dulić V, Kaufmann WK, Wilson SJ, Tisty TD, Lees E, Harper JW, Elledge SJ and Reed SI: p53-dependent inhibition of cyclin-dependent kinase activities in human fibroblasts during radiation-induced G1 arrest. Cell. 76:1013–1023. 1994. View Article : Google Scholar
20	MacLachlan TK, Sang N and Giordano A: Cyclins, cyclin-dependent kinases and cdk inhibitors: implications in cell cycle control and cancer. Crit Rev Eukaryot Gene. 5:127–156. 1995. View Article : Google Scholar
21	Murray A: Cell cycle checkpoints. Curr Opin Cell Biol. 6:872–876. 1994. View Article : Google Scholar : PubMed/NCBI
22	Parthiban P, Aridoss G, Rathika P, Ramkumar V and Kabilan S: Synthesis, stereochemistry and antimicrobial studies of novel oxime ethers of aza/diazabicycles. Bioorg Med Chem Lett. 19:6981–6985. 2009. View Article : Google Scholar : PubMed/NCBI
23	Haridas V, Rajgokul KS, Sadanandan S, Agrawal T, Sharvani V, Gopalakrishna M, Bijesh M, Kumawat KL, Basu A and Medigeshi GR: Bispidine-amino acid conjugates act as a novel scaffold for the design of antivirals that block Japanese encephalitis virus replication. PLoS Negl Trop Dis. 7:20052013. View Article : Google Scholar
24	Tominaga H, Ishiyama M, Ohseto F, Sasamoto K, Hamamoto T, Suzuki K and Watanabe M: A water-soluble tetrazolium salt useful for colorimetric cell viability assay. Anal Commun. 36:47–50. 1999. View Article : Google Scholar
25	Lin SY, Liu JD, Chang HC, Yeh SD, Lin CH and Lee WS: Magnolol suppresses proliferation of cultured human colon and liver cancer cells by inhibiting DNA synthesis and activating apoptosis. J Cell Biochem. 84:532–544. 2002. View Article : Google Scholar : PubMed/NCBI
26	Bobyleva V, Pazienza TL, Maseroli R, Tomasi A, Salvioli S, Cossarizza A, Franceschi C and Skulachev VP: Decrease in mitochondrial energy coupling by thyroid hormones: a physiological effect rather than a pathological hyperthyroidism consequence. FEBS Lett. 430:409–413. 1998. View Article : Google Scholar : PubMed/NCBI
27	Collins KL and Kelly TJ: Effects of T antigen and replication protein A on the initiation of DNA synthesis by DNA polymerase alpha-primase. Mol Cell Biol. 11:2108–2115. 1991.PubMed/NCBI
28	Tang XQ, Feng JQ, Chen J, Chen PX, Zhi JL, Cui Y, Guo RX and Yu HM: Protection of oxidative preconditioning against apoptosis induced by H2O2 in PC12 cells: mechanisms via MMP, ROS, and Bcl-2. Brain Res. 1057:57–64. 2005. View Article : Google Scholar : PubMed/NCBI
29	Shiff SJ, Qiao L, Tsai LL and Rigas B: Sulindac sulfide, an aspirin-like compound, inhibits proliferation, causes cell cycle quiescence, and induces apoptosis in HT-29 colon adenocarcinoma cells. J Clin Invest. 96:491–503. 1995. View Article : Google Scholar : PubMed/NCBI
30	Boehme SA and Lenardo MJ: Propriocidal apoptosis of mature T lymphocytes occurs at S phase of the cell cycle. Eur J Immunol. 23:1552–1560. 1993. View Article : Google Scholar : PubMed/NCBI
31	Zhang HS, Gavin M, Dahiya A, Postigo AA, Ma D, Luo RX, Harbour JW and Dean DC: Exit from G1 and S phase of the cell cycle is regulated by repressor complexes containing HDAC-Rb-hSWI/SNF and Rb-hSWI/SNF. Cell. 101:79–89. 2000. View Article : Google Scholar : PubMed/NCBI
32	Thomas A, El Rouby S, Reed JC, Krajewski S, Silber R, Potmesil M and Newcomb EW: Drug-induced apoptosis in B-cell chronic lymphocytic leukemia: relationship between p53 gene mutation and bcl-2/bax proteins in drug resistance. Oncogene. 12:1055–1062. 1996.PubMed/NCBI
33	Yin X, Oltvai ZN and Korsmeyer SJ: BH1 and BH2 domains of Bcl-2 are required for inhibition of apoptosis and heterodimerization with Bax. Nature. 369:321–323. 1994. View Article : Google Scholar : PubMed/NCBI
34	Miyashita T, Krajewski S, Krajewska M, Wang HG, Lin H, Liebermann DA, Hoffman B and Reed JC: Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene. 9:1799–1805. 1994.PubMed/NCBI
35	Fulda S and Debatin KM: Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene. 25:4798–4811. 2006. View Article : Google Scholar : PubMed/NCBI
36	Saitoh M, Nagai K, Nakagawa K, Yamamura T, Yamamoto S and Nishizaki T: Adenosine induces apoptosis in the human gastric cancer cells via an intrinsic pathway relevant to activation of AMP-activated protein kinase. Biochem Pharmacol. 67:2005–2011. 2004. View Article : Google Scholar : PubMed/NCBI