The endogenous oxindole isatin induces apoptosis of MCF‑7 breast cancer cells through a mitochondrial pathway

Ma,Zhongliang; Hou,Lin; Jiang,Yuhong; Chen,Yanpin; Song,Jinlian

doi:10.3892/or.2014.3426

The endogenous oxindole isatin induces apoptosis of MCF‑7 breast cancer cells through a mitochondrial pathway

Authors:
- Zhongliang Ma
- Lin Hou
- Yuhong Jiang
- Yanpin Chen
- Jinlian Song
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Affiliations: Department of Breast Surgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266000, P.R. China, Department of Biochemistry, Medical College, Qingdao University, Qingdao, Shandong 266021, P.R. China, Department of Laboratory, Women and Children's Hospital of Qingdao, Qingdao, Shandong 266011, P.R. China
Published online on: August 20, 2014 https://doi.org/10.3892/or.2014.3426
Pages: 2111-2117

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Abstract

Isatin is an endogenous indole in mammalian tissues and fluids that is expected to have antitumor effects in human breast cancer cells. Human breast cancer cells (MCF-7) were exposed to isatin at various concentrations (0, 50, 100, 200 µmol/l) for 48 h. Apoptotic features were demonstrated by nuclei staining with Hoechst 33258 and flow cytometry. Bcl-2 and Bax mRNA were analyzed via reverse transcription-polymerase chain reaction. Bcl-2, Bax, the inhibitor of caspase-activated DNase (ICAD), and cytochrome c protein were analyzed by western blot analysis. Apoptosis, caspase-9 and -3 activation and mitochondrial depolarization were assayed by flow cytometry. The results showed that isatin induced apoptosis of MCF-7 cells. Furthermore, Bcl-2 expression was decreased and the ratio of Bcl-2 to Bax was significantly decreased by isatin. The mitochondrial transmembrane potential was markedly decreased and the release of cytochrome c into the cytosol was elevated following treatment with isatin. At the same time, caspase-9 and -3 were stimulated, followed by the degradation of ICAD, a caspase-3 substrate.

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1	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar
2	DeSantis C, Siegel R, Bandi P and Jemal A: Breast cancer statistics, 2011. CA Cancer J Clin. 61:409–418. 2011. View Article : Google Scholar
3	Smith NZ: Treating metastatic breast cancer with systemic chemotherapies: current trends and future perspectives. Clin J Oncol Nurs. 16:E33–E43. 2012. View Article : Google Scholar : PubMed/NCBI
4	Gonzalez-Angulo AM, Morales-Vasquez F and Hortobagyi GN: Overview of resistance to systemic therapy in patients with breast cancer. Adv Exp Med Biol. 608:1–22. 2007. View Article : Google Scholar : PubMed/NCBI
5	Ramachandran S: Synthesis and antimicrobial evaluation of some novel schiff and mannich bases of isatin derivatives. Int J Res Pharm Chem. 1:289–294. 2011.
6	Medvedev A, Igosheva N, Crumeyrolle-Arias M and Glover V: Isatin: role in stress and anxiety. Stress. 8:175–183. 2005. View Article : Google Scholar : PubMed/NCBI
7	Pandeya SN, Smitha S, Jyoti M and Sridhar SK: Biological activities of isatin and its derivatives. Acta Pharm. 55:27–46. 2005.PubMed/NCBI
8	Medvedev A, Buneeva O and Glover V: Biological targets for isatin and its analogues: implications for therapy. Biologics. 1:151–162. 2007.PubMed/NCBI
9	Vine KL, Matesic L, Locke JM, Ranson M and Skropeta D: Cytotoxic and anticancer activities of isatin and its derivatives: a comprehensive review from 2000–2008. Anticancer Agents Med Chem. 9:397–414. 2009.PubMed/NCBI
10	Cane A, Tournaire MC, Barritault D and Crumeyrolle-Arias M: The endogenous oxindoles 5-hydroxyoxindole and isatin are antiproliferative and proapoptotic. Biochem Biophys Res Commun. 276:379–384. 2000. View Article : Google Scholar : PubMed/NCBI
11	Sun L, Liang C, Shirazian S, Zhou Y, Miller T, Cui J, Fukuda JY, et al: Discovery of 5-[5-fluoro-2-oxo-1,2-dihydroindol-(3Z)-ylidenemethyl]-2,4-dimethyl-1H-pyrrole-3-carboxylic acid (2-diethylaminoethyl) amide, a novel tyrosine kinase inhibitor targeting vascular endothelial and platelet-derived growth factor receptor tyrosine kinase. J Med Chem. 46:1116–1119. 2003.
12	Polychronopoulos P, Magiatis P, Skaltsounis AL, Myrianthopoulos V, Mikros E, Tarricone A, Musacchio A, et al: Structural basis for the synthesis of indirubins as potent and selective inhibitors of glycogen synthase kinase-3 and cyclin-dependent kinases. J Med Chem. 47:935–946. 2004. View Article : Google Scholar : PubMed/NCBI
13	Krishnegowda G, Prakasha Gowda AS, Tagaram HR, Carroll KF, Irby RB, Sharma AK and Amin S: Synthesis and biological evaluation of a novel class of isatin analogs as dual inhibitors of tubulin polymerization and Akt pathway. Bioorg Med Chem. 19:6006–6014. 2011. View Article : Google Scholar : PubMed/NCBI
14	Wang C and Youle RJ: The role of mitochondria in apoptosis. Annu Rev Genet. 43:95–118. 2009. View Article : Google Scholar
15	Volkmann N, Marassi FM, Newmeyer DD and Hanein D: The rheostat in the membrane: BCL-2 family proteins and apoptosis. Cell Death Differ. 21:206–215. 2014. View Article : Google Scholar : PubMed/NCBI
16	Cory S, Huang DC and Adams JM: The Bcl-2 family: roles in cell survival and oncogenesis. Oncogene. 24:8590–8607. 2003. View Article : Google Scholar : PubMed/NCBI
17	Cottet-Rousselle C, Ronot X, Leverve X and Mayol JF: Cytometric assessment of mitochondria using fluorescent probes. Cytometry A. 79:405–425. 2011. View Article : Google Scholar : PubMed/NCBI
18	Pakravan P, Kashanian S, Khodaei MM and Harding FJ: Biochemical and pharmacological characterization of isatin and its derivatives: from structure to activity. Pharmacol Rep. 65:313–335. 2013. View Article : Google Scholar : PubMed/NCBI
19	Premanathan M, Radhakrishnan S, Kulangiappar K, Singaravelu G, Thirumalaiarasu V, Sivakumar T and Kathiresan K: Antioxidant & anticancer activities of isatin (1H-indole-2,3-dione), isolated from the flowers of Couroupita guianensis Aubl. Indian J Med Res. 136:822–826. 2012.
20	de Cândido-Bacani PM, Mori MP, Calvo TR, Vilegas W, Varanda EA and Cólus IM: In vitro assessment of the cytotoxic, apoptotic, and mutagenic potentials of isatin. J Toxicol Environ Health A. 76:354–362. 2013.PubMed/NCBI
21	Ashkenazi A and Dixit VM: Death receptors: signaling and modulation. Science. 281:1305–1308. 1998. View Article : Google Scholar : PubMed/NCBI
22	Reed JC: Double identity for proteins of the Bcl-2 family. Nature. 387:773–776. 1997. View Article : Google Scholar : PubMed/NCBI
23	Bedikian AY, Millward M, Pehamberger H, Conry R, Gore M, Trefzer U, Pavlick AC, et al: Bcl-2 antisense (oblimersen sodium) plus dacarbazine in patients with advanced melanoma: the oblimersen melanoma study group. J Clin Oncol. 24:4738–4745. 2006. View Article : Google Scholar : PubMed/NCBI
24	Indran IR, Tufo G, Pervaiz S and Brenner C: Recent advances in apoptosis, mitochondria and drug resistance in cancer cells. Biochim Biophys Acta. 1807:735–745. 2011. View Article : Google Scholar : PubMed/NCBI
25	Hengartner MO: The biochemistry of apoptosis. Nature. 407:770–776. 2000. View Article : Google Scholar : PubMed/NCBI
26	Kuranaga E: Caspase signaling in animal development. Dev Growth Differ. 53:137–148. 2011. View Article : Google Scholar
27	Yoshida A, Pommier Y and Ueda T: Endonuclease activation and chromosomal DNA fragmentation during apoptosis in leukemia cells. Int J Hematol. 84:31–37. 2006. View Article : Google Scholar : PubMed/NCBI