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

Jeong,Hyunjin; Shin,Yumi; Kim,Dong‑Kyoo

doi:10.3892/or.2018.6404

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

Authors:
- Hyunjin Jeong
- Yumi Shin
- Dong‑Kyoo Kim
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Affiliations: Department of Biomedicinal Chemistry and Institute of Basic Science, Inje University, Gimhae, Gyeongsang 621‑749, Republic of Korea
Published online on: April 27, 2018 https://doi.org/10.3892/or.2018.6404
Pages: 545-553

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Abstract

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.

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1	Parkin DM: Global cancer statistics in the year 2000. Lancet Oncol. 2:533–543. 2001. View Article : Google Scholar : PubMed/NCBI
2	World Health Organization, . Control of cancer of the cervix uteri: Review article based on a report of a WHO meeting, November 1985, Geneva. Bull World Health Organ. 64:607–618. 1986.PubMed/NCBI
3	Yaoxian W, Hui Y, Yunyan Z, Yanqin L, Xin G and Xiaoke W: Emodin induces apoptosis of human cervical cancer HeLa cells via intrinsic mitochondrial and extrinsic death receptor pathway. Cancer Cell Int. 13:712013. View Article : Google Scholar : PubMed/NCBI
4	Reed JC: Apoptosis-regulating proteins as targets for drug discovery. Trends Mol Med. 7:314–319. 2001. View Article : Google Scholar : PubMed/NCBI
5	Earnshaw WC: Nuclear changes in apoptosis. Curr Opin Cell Biol. 7:337–343. 1995. View Article : Google Scholar : PubMed/NCBI
6	Degterev A, Boyce M and Yuan J: A decade of caspases. Oncogene. 22:8543–8567. 2003. View Article : Google Scholar : PubMed/NCBI
7	Ziegler DS and Kung AL: Therapeutic targeting of apoptosis pathways in cancer. Curr Opin Oncol. 20:97–103. 2008. View Article : Google Scholar : PubMed/NCBI
8	Ehrlich E: Regulation of BAX mediated apoptosis by BCL2 family members. SABiosciences. 2011.https://www.qiagen.com/fi/spotlight-pages/newsletters-and-magazines/articles/reviews-online-apoptosis/
9	Reed JC and Green DR: Remodeling for demolition: Changes in mitochondrial ultrastructure during apoptosis. Mol Cell. 9:1–3. 2002. View Article : Google Scholar : PubMed/NCBI
10	Wang C and Youle RJ: The role of mitochondria in apoptosis. Annu Rev Genet. 43:95–118. 2009. View Article : Google Scholar : PubMed/NCBI
11	Ghobrial IM, Witzig TE and Adjei AA: Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin. 55:178–194. 2005. View Article : Google Scholar : PubMed/NCBI
12	Marzo I, Brenner C, Zamzami N, Susin SA, Beutner G, Brdiczka D, Rémy R, Xie ZH, Reed JC and Kroemer G: The permeability transition pore complex: A target for apoptosis regulation by caspases and bcl-2-related proteins. J Exp Med. 187:1261–1271. 1998. View Article : Google Scholar : PubMed/NCBI
13	Kuwana T and Newmeyer DD: Bcl-2-family proteins and the role of mitochondria in apoptosis. Curr Opin Cell Biol. 15:691–699. 2003. View Article : Google Scholar : PubMed/NCBI
14	Gross A, McDonnell JM and Korsmeyer SJ: BCL-2 family members and the mitochondria in apoptosis. Genes Dev. 13:1899–1911. 1999. View Article : Google Scholar : PubMed/NCBI
15	Zhang X, Zhao J, Kang S, Yi M, You S, Shin DS and Kim DK: A novel cromakalim analogue induces cell cycle arrest and apoptosis in human cervical carcinoma HeLa cells through the caspase- and mitochondria-dependent pathway. Int J Oncol. 39:1609–1617. 2011.PubMed/NCBI
16	LeBlanc H, Lawrence D, Varfolomeev E, Totpal K, Morlan J, Schow P, Fong S, Schwall R, Sinicropi D and Ashkenazi A: Tumor-cell resistance to death receptor - induced apoptosis through mutational inactivation of the proapoptotic Bcl-2 homolog Bax. Nat Med. 8:274–281. 2002. View Article : Google Scholar : PubMed/NCBI
17	Inal ME, Kanbak G and Sunal E: Antioxidant enzyme activities and malondialdehyde levels related to aging. Clin Chim Acta. 305:75–80. 2001. View Article : Google Scholar : PubMed/NCBI
18	Le Bras M, Clément MV, Pervaiz S and Brenner C: Reactive oxygen species and the mitochondrial signaling pathway of cell death. Histol Histopathol. 20:205–219. 2005.PubMed/NCBI
19	Fleury C, Mignotte B and Vayssière JL: Mitochondrial reactive oxygen species in cell death signaling. Biochimie. 84:131–141. 2002. View Article : Google Scholar : PubMed/NCBI
20	Simon HU, Haj-Yehia A and Levi-Schaffer F: Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis. 5:415–418. 2000. View Article : Google Scholar : PubMed/NCBI
21	Chan WH, Wu CC and Yu JS: Curcumin inhibits UV irradiation-induced oxidative stress and apoptotic biochemical changes in human epidermoid carcinoma A431 cells. J Cell Biochem. 90:327–338. 2003. View Article : Google Scholar : PubMed/NCBI
22	Jiang CP, Ding H, Shi DH, Wang YR, Li EG and Wu JH: Pro-apoptotic effects of tectorigenin on human hepatocellular carcinoma HepG2 cells. World J Gastroenterol. 18:1753–1764. 2012. View Article : Google Scholar : PubMed/NCBI
23	Sherr CJ: Cancer cell cycles. Science. 274:1672–1677. 1996. View Article : Google Scholar : PubMed/NCBI
24	Hartwell LH and Kastan MB: Cell cycle control and cancer. Science. 266:1821–1828. 1994. View Article : Google Scholar : PubMed/NCBI
25	Dictor M, Ehinger M, Mertens F, Akervall J and Wennerberg J: Abnormal cell cycle regulation in malignancy. Am J Clin Pathol. 112 Suppl 1:S40–S52. 1999.PubMed/NCBI
26	Bonelli P, Tuccillo FM, Borrelli A, Schiattarella A and Buonaguro FM: CDK/CCN and CDKI alterations for cancer prognosis and therapeutic predictivity. BioMed Res Int. 2014:3610202014. View Article : Google Scholar : PubMed/NCBI
27	Lu YJ, Yang SH, Chien CM, Lin YH, Hu XW, Wu ZZ, Wu MJ and Lin SR: Induction of G2/M phase arrest and apoptosis by a novel enediyne derivative, THDB, in chronic myeloid leukemia (HL-60) cells. Toxicol In Vitro. 21:90–98. 2007. View Article : Google Scholar : PubMed/NCBI
28	Goffin E, Lamoral-Theys D, Tajeddine N, de Tullio P, Mondin L, Lefranc F, Gailly P, Rogister B, Kiss R and Pirotte B: 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. Eur J Med Chem. 54:834–844. 2012. View Article : Google Scholar : PubMed/NCBI
29	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
30	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
31	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
32	Qi F, Li A, Zhao L, Xu H, Inagaki Y, Wang D, Cui X, Gao B, Kokudo N, Nakata M, et al: Cinobufacini, an aqueous extract from Bufo bufo gargarizans Cantor, induces apoptosis through a mitochondria-mediated pathway in human hepatocellular carcinoma cells. J Ethnopharmacol. 128:654–661. 2010. View Article : Google Scholar : PubMed/NCBI
33	Park MT, Kim MJ, Kang YH, Choi SY, Lee JH, Choi JA, Kang CM, Cho CK, Kang S, Bae S, et al: Phytosphingosine in combination with ionizing radiation enhances apoptotic cell death in radiation-resistant cancer cells through ROS-dependent and -independent AIF release. Blood. 105:1724–1733. 2005. View Article : Google Scholar : PubMed/NCBI
34	Lee S, Christakos S and Small MB: Apoptosis and signal transduction: Clues to a molecular mechanism. Curr Opin Cell Biol. 5:286–291. 1993. View Article : Google Scholar : PubMed/NCBI
35	Alenzi FQ: Links between apoptosis, proliferation and the cell cycle. Br J Biomed Sci. 61:99–102. 2004. View Article : Google Scholar : PubMed/NCBI
36	Cao W, Li XQ, Wang X, Fan HT, Zhang XN, Hou Y, Liu SB and Mei QB: A novel polysaccharide, isolated from Angelica sinensis (Oliv.) Diels induces the apoptosis of cervical cancer HeLa cells through an intrinsic apoptotic pathway. Phytomedicine. 17:598–605. 2010. View Article : Google Scholar : PubMed/NCBI
37	Chen S, Cheng AC, Wang MS and Peng X: Detection of apoptosis induced by new type gosling viral enteritis virus in vitro through fluorescein annexin V-FITC/PI double labeling. World J Gastroenterol. 14:2174–2178. 2008. View Article : Google Scholar : PubMed/NCBI
38	Reed JC, Jurgensmeier JM and Matsuyama S: Bcl-2 family proteins and mitochondria. Biochim Biophys Acta. 1366:127–137. 1998. View Article : Google Scholar : PubMed/NCBI
39	Oltvai ZN, Milliman CL and Korsmeyer SJ: Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell. 74:609–619. 1993. View Article : Google Scholar : PubMed/NCBI
40	Lee YS, Sayeed MM and Wurster RD: In vitro antitumor activity of cromakalim in human brain tumor cells. Pharmacology. 49:69–74. 1994. View Article : Google Scholar : PubMed/NCBI
41	Kiechle FL and Zhang X: Apoptosis: Biochemical aspects and clinical implications. Clin Chim Acta. 326:27–45. 2002. View Article : Google Scholar : PubMed/NCBI
42	Otsuki Y, Li Z and Shibata MA: Apoptotic detection methods - from morphology to gene. Prog Histochem Cytochem. 38:275–339. 2003. View Article : Google Scholar : PubMed/NCBI
43	Sánchez I and Dynlacht BD: New insights into cyclins, CDKs, and cell cycle control. Semin Cell Dev Biol. 16:311–321. 2005. View Article : Google Scholar : PubMed/NCBI
44	Smith DM, Gao G, Zhang X, Wang G and Dou QP: Regulation of tumor cell apoptotic sensitivity during the cell cycle (Review). Int J Mol Med. 6:503–507. 2000.PubMed/NCBI
45	Zhang Y, Ahn EY, Jiang Y, Kim DK, Kang SG, Wu C, Kang SW, Park JS, Son BW and Jung JH: 3-Chloro-2,5-dihydroxybenzyl alcohol activates human cervical carcinoma HeLa cell apoptosis by inducing DNA damage. Int J Oncol. 31:1317–1323. 2007.PubMed/NCBI
46	Hwang J, Yi M, Zhang X, Xu Y, Jung JH and Kim DK: Cytochalasin B induces apoptosis through the mitochondrial apoptotic pathway in HeLa human cervical carcinoma cells. Oncol Rep. 30:1929–1935. 2013. View Article : Google Scholar : PubMed/NCBI
47	Qi F, Inagaki Y, Gao B, Cui X, Xu H, Kokudo N, Li A and Tang W: Bufalin and cinobufagin induce apoptosis of human hepatocellular carcinoma cells via Fas- and mitochondria-mediated pathways. Cancer Sci. 102:951–958. 2011. View Article : Google Scholar : PubMed/NCBI