Anticancer effects of a novel chroman analog in HeLa cells are associated with G2‑phase arrest and mitochondrial‑mediated apoptosis.

In the present study, the anticancer activity of 1‑[(3S,4R)‑2,2‑dimethyl‑3‑oxo‑4‑(2‑piperidonyl)chroman‑6‑yl]‑3‑phenylurea (S32) was investigated by testing its effect in vitro on the growth of HeLa cells. First, we showed that the IC50 value of S32 was ~70 µM by using WST‑8 assay, and that it significantly inhibited the proliferation and viability of HeLa cells in a dose‑dependent manner after 48 h. Morphological changes in apoptotic cells included cellular shrinkage and nuclear condensation. The results of [3H]‑thymidine incorporation and flow cytometric analysis indicated that S32 induced inhibition of DNA replication and G2‑phase cell cycle arrest. Moreover, S32 induced the levels of reactive oxygen species (ROS) and decreased the mitochondrial membrane potential (MMP) in a time‑dependent manner. Using Annexin V‑FITC/propidium iodide (PI) dual staining assay, we found that S32 noticeably increased early apoptosis in HeLa cells in a time‑dependent manner. The result of western blot analysis showed that the apoptotic induction was associated with an increase in Bax levels and a decrease in Bcl‑2 levels, which led to activation of caspase‑8, ‑9 and ‑3. Taken together, our findings demonstrated that S32 induces mitochondrial‑mediated apoptosis in HeLa cells and suggest that S32 has potential as an anticancer drug.

[1]  Samir Guglani.  Death , 1890, The Lancet.

[2]  F. Buonaguro,et al.  CDK/CCN and CDKI Alterations for Cancer Prognosis and Therapeutic Predictivity , 2014, BioMed research international.

[3]  Xin Zhang,et al.  Cytochalasin B induces apoptosis through the mitochondrial apoptotic pathway in HeLa human cervical carcinoma cells. , 2013, Oncology reports.

[4]  L. Yanqin,et al.  Emodin induces apoptosis of human cervical cancer hela cells via intrinsic mitochondrial and extrinsic death receptor pathway , 2013, Cancer Cell International.

[5]  R. Kiss,et al.  N-Aryl-N'-(chroman-4-yl)ureas and thioureas display in vitro anticancer activity and selectivity on apoptosis-resistant glioblastoma cells: screening, synthesis of simplified derivatives, and structure-activity relationship analysis. , 2012, European journal of medicinal chemistry.

[6]  Junhua Wu,et al.  Pro-apoptotic effects of tectorigenin on human hepatocellular carcinoma HepG2 cells. , 2012, World journal of gastroenterology.

[7]  Xin Zhang,et al.  A novel cromakalim analogue induces cell cycle arrest and apoptosis in human cervical carcinoma HeLa cells through the caspase- and mitochondria-dependent pathway. , 2011, International journal of oncology.

[8]  W. Cao,et al.  A novel polysaccharide, isolated from Angelica sinensis (Oliv.) Diels induces the apoptosis of cervical cancer HeLa cells through an intrinsic apoptotic pathway. , 2010, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[9]  Wei Tang,et al.  Cinobufacini, an aqueous extract from Bufo bufo gargarizans Cantor, induces apoptosis through a mitochondria-mediated pathway in human hepatocellular carcinoma cells. , 2010, Journal of ethnopharmacology.

[10]  A. Cheng,et al.  Detection of apoptosis induced by new type gosling viral enteritis virus in vitro through fluorescein annexin V-FITC/PI double labeling. , 2008, World journal of gastroenterology.

[11]  Seon-Yong Jeong,et al.  The role of mitochondria in apoptosis. , 2008, BMB reports.

[12]  David S Ziegler,et al.  Therapeutic targeting of apoptosis pathways in cancer , 2008, Current opinion in oncology.

[13]  Chunfu Wu,et al.  3-Chloro-2,5-dihydroxybenzyl alcohol activates human cervical carcinoma HeLa cell apoptosis by inducing DNA damage. , 2007, International journal of oncology.

[14]  Sheng-Huei Yang,et al.  Induction of G2/M phase arrest and apoptosis by a novel enediyne derivative, THDB, in chronic myeloid leukemia (HL-60) cells. , 2007, Toxicology in vitro : an international journal published in association with BIBRA.

[15]  Brian David Dynlacht,et al.  New insights into cyclins, CDKs, and cell cycle control. , 2005, Seminars in cell & developmental biology.

[16]  Jung-A Choi,et al.  Phytosphingosine in combination with ionizing radiation enhances apoptotic cell death in radiation-resistant cancer cells through ROS-dependent and -independent AIF release. , 2005, Blood.

[17]  S. Pervaiz,et al.  Reactive oxygen species and the mitochondrial signaling pathway of cell death. , 2005, Histology and histopathology.

[18]  S. Fulda,et al.  Targeting apoptosis pathways in cancer therapy. , 2004, Current cancer drug targets.

[19]  F. Alenzi,et al.  Links between apoptosis, proliferation and the cell cycle , 2004, British journal of biomedical science.

[20]  F. Levi-Schaffer,et al.  Role of reactive oxygen species (ROS) in apoptosis induction , 2000, Apoptosis.

[21]  T. Kuwana,et al.  Bcl-2-family proteins and the role of mitochondria in apoptosis. , 2003, Current opinion in cell biology.

[22]  Alexei Degterev,et al.  A decade of caspases , 2003, Oncogene.

[23]  Jau-Song Yu,et al.  Curcumin inhibits UV irradiation‐induced oxidative stress and apoptotic biochemical changes in human epidermoid carcinoma A431 cells , 2003, Journal of cellular biochemistry.

[24]  M. Shibata,et al.  Apoptotic detection methods--from morphology to gene. , 2003, Progress in histochemistry and cytochemistry.

[25]  F. Kiechle,et al.  Apoptosis: biochemical aspects and clinical implications. , 2002, Clinica chimica acta; international journal of clinical chemistry.

[26]  Peter Schow,et al.  Tumor-cell resistance to death receptor–induced apoptosis through mutational inactivation of the proapoptotic Bcl-2 homolog Bax , 2002, Nature Medicine.

[27]  B. Mignotte,et al.  Mitochondrial reactive oxygen species in cell death signaling. , 2002, Biochimie.

[28]  Chien-Huang Lin,et al.  Magnolol suppresses proliferation of cultured human colon and liver cancer cells by inhibiting DNA synthesis and activating apoptosis , 2002, Journal of cellular biochemistry.

[29]  D. Green,et al.  Remodeling for demolition: changes in mitochondrial ultrastructure during apoptosis. , 2002, Molecular cell.

[30]  D. Parkin,et al.  Global cancer statistics in the year 2000. , 2001, The Lancet. Oncology.

[31]  J C Reed,et al.  Apoptosis-regulating proteins as targets for drug discovery. , 2001, Trends in molecular medicine.

[32]  M. Inal,et al.  Antioxidant enzyme activities and malondialdehyde levels related to aging. , 2001, Clinica chimica acta; international journal of clinical chemistry.

[33]  D. Smith,et al.  Regulation of tumor cell apoptotic sensitivity during the cell cycle (Review). , 2000, International journal of molecular medicine.

[34]  S. Korsmeyer,et al.  BCL-2 family members and the mitochondria in apoptosis. , 1999, Genes & development.

[35]  F. Mertens,et al.  Abnormal cell cycle regulation in malignancy. , 1999, American journal of clinical pathology.

[36]  T. Hamamoto,et al.  A water-soluble tetrazolium salt useful for colorimetric cell viability assay , 1999 .

[37]  John Calvin Reed,et al.  Bcl-2 family proteins and mitochondria. , 1998, Biochimica et biophysica acta.

[38]  C. Franceschi,et al.  Decrease in mitochondrial energy coupling by thyroid hormones: a physiological effect rather than a pathological hyperthyroidism consequence , 1998, FEBS letters.

[39]  G. Kroemer,et al.  The Permeability Transition Pore Complex: A Target for Apoptosis Regulation by Caspases and Bcl-2–related Proteins , 1998, The Journal of experimental medicine.

[40]  C. Sherr.  Cancer Cell Cycles , 1996, Science.

[41]  W. Earnshaw,et al.  Nuclear changes in apoptosis. , 1995, Current opinion in cell biology.

[42]  K W Kohn,et al.  Cell cycle control and cancer chemotherapy , 1994, Journal of cellular biochemistry.

[43]  Y. Lee,et al.  In vitro antitumor activity of cromakalim in human brain tumor cells. , 1994, Pharmacology.

[44]  S. Christakos,et al.  Apoptosis and signal transduction: clues to a molecular mechanism. , 1993, Current opinion in cell biology.

[45]  B. Stepowski.  [Control of cancer of the cervix uteri]. , 1953, Ginekologia polska.