The radiation-sensitizing effect of flavopiridol in the esophageal cancer cell line Eca109

Yao,Yuan; Shi,Jingbin; Zhang,Zhuo; Zhang,Feng; Ma,Ruilan; Zhao,Yan

doi:10.3892/ol.2013.1291

The radiation-sensitizing effect of flavopiridol in the esophageal cancer cell line Eca109

Authors:
- Yuan Yao
- Jingbin Shi
- Zhuo Zhang
- Feng Zhang
- Ruilan Ma
- Yan Zhao
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Affiliations: Department of Radiology, The Second Hospital of Dalian Medical University, Dalian 116000, P.R. China
Published online on: April 4, 2013 https://doi.org/10.3892/ol.2013.1291
Pages: 1872-1876

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Abstract

Flavopiridol is a cyclin-dependent kinase inhibitor. It has shown an antitumor effect against several cancers. In the present study, the radiation-sensitizing effect of flavopiridol was investigated in an esophageal squamous carcinoma cell line, Eca109. The growth inhibitory rate of Eca109 with flavopiridol was determined using the MTT and the radiosensitizing rate using clonogenic survival assays. The cell cycle distribution and the rate of apoptosis were measured using flow cytometry. The proteins cyclin D1, ERK/pERK, caspase-3, Bax and Bcl-2 were detected using western blot analysis to elucidate the mechanism of the radiosensitization effect. MTT assay showed that flavopiridol inhibited the survival rate of Eca109 cells and the effect was dose-dependent. Its IC50 was 193.3 nmol/l. The result of the clonogenic survival revealed that flavopiridol enhanced the radiosensitivity of Eca109 cells and the sensitization enhancement ratio (SER) was 1.194 at 0.2xIC50. Moreover, we detected that the cells treated with flavorpiridol were arrested at the G2/M phase and the apoptosis caused by radiation was increased. In addition, the proteins caspase-3 and Bax in cells treated with flavopiridol were upregulated, while cyclin D1 and Bcl-2 were downregulated. In conclusion, flavopiridol may enhance the radiosensitivity of Eca109 cells and the radiosensitizing effect of flavopiridol may be mediated by decreasing the levels of the cyclin D1 protein, thus increasing the percentage of cells at G2/M phase.

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1	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: GLOBOCAN 2008 v1.2, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 10 (Internet). International Agency for Research on Cancer; Lyon, France: pp. 2010http://globocan.iarc.fr. Accessed March 15, 2012.
2	Brown LM and Devesa SS: Epidemiologic trends in esophagealand gastric cancer in the United States. Surg Oncol Clin N Am. 11:235–256. 2002. View Article : Google Scholar : PubMed/NCBI
3	Pawlik TM and Keyomarsi K: Role of cell cycle in mediating sensitivity to radiotherapy. Int J Radiat Oncol Biol Phys. 4:928–942. 2004. View Article : Google Scholar : PubMed/NCBI
4	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 Expr. 5:127–156. 1995. View Article : Google Scholar : PubMed/NCBI
5	Zafonte BT, Hulit J, Amanatullah DF, Albanese C, Wang C, Rosen E, Reutens A, Sparano JA, Lisanti MP and Pestell RG: Cell-cycle dysregulation in breast cancer therapies targeting the cell cycle. Front Biosci. 5:938–961. 2000. View Article : Google Scholar : PubMed/NCBI
6	Sedlacek HH: Mechanisms of action of flavopiridol. Crit Rev Oncol Hematol. 38:139–170. 2001. View Article : Google Scholar
7	Monga M and Sausville EA: Developmental therapeutics program at the NCI: molecular target and drug discovery process. Leukemia. 16:520–526. 2002. View Article : Google Scholar : PubMed/NCBI
8	Senderowicz AM and Sausville: Preclinical and clinical development of cyclin-dependent kinase modulators. J Natl Cancer Inst. 92:376–387. 2000. View Article : Google Scholar : PubMed/NCBI
9	Kaur G, Stetler-Stevenson M, Sebers S, Worland P, Sedlacek H, Myers C, Czech J, Naik R and Sausville E: Growth inhibition with reversible cell cycle arrest of carcinoma cells by flavone L86-8275. JNCI J Natl Cancer Inst. 84:1736–1740. 1992. View Article : Google Scholar : PubMed/NCBI
10	Camphausen K, Brady K, Burgan W, Carra M, Russell J, Bull E and Tofilon P: Flavopiridol enhances human tumor cell radio-sensitivity and prolongs expression of gammaH2AX foci. Mol Cancer Ther. 3:409–416. 2004.PubMed/NCBI
11	Byrd JC, Shinn C, Waselenko JK, Fuchs EJ, Lehman TA, Nguyen PL, Flinn IW, Diehl LF, Sausville E and Grever MR: Flavopiridol induces apoptosis in chronic lymphocytic leukemia cells via activation of caspase-3 without evidence of bcl-2 modulation or dependence on functional p53. Blood. 92:3804–3816. 2008.PubMed/NCBI
12	Patel V, Senderowicz AM, Pinto D, Igishi T, Raffeld M, Quintanilla-Martinez L, Ensley JF, Sausville EA and Gutkind JS: Flavopiridol, a novel cyclin-dependent kinase inhibitor, suppresses the growth of head and neck squamous cell carcinomas by inducing apoptosis. J Clin Invest. 102:1674–1681. 1998. View Article : Google Scholar : PubMed/NCBI
13	Li Y, Bhuiyan M and Sarkar FH: Induction of apoptosis and inhibition of c-erbB-2 in MDA-MB-435 cells by genistein. Int J Oncol. 15:525–533. 1999.PubMed/NCBI
14	Fredersdorf S, Burns J, Milne AM, Packham G, Fallis L, Gillett CE, Royds JA, Peston D, Hall PA, Hanby AM, Barnes DM, Shousha S, O’Hare MJ and Lu X: High level expression of p27kip1 and cyclin D1 in some human breast cancer cells: Inverse correlation between the expression of p27kip1 and degree of malignancy in human breast and colorectal cancers. Proc Natl Acad Sci USA. 94:6380–6385. 1997. View Article : Google Scholar : PubMed/NCBI
15	Gansauge S, Gansauge F, Ramadani M, Stobbe H, Rau B, Harada N and Beger HG: Overexpression of cyclin D1 in human pancreatic carcinoma is associated with poor prognosis. Cancer Res. 57:1634–1637. 1997.PubMed/NCBI
16	Adélaide J, Monges G, Dérdérian C, Seitz JF and Birnbaum D: Esophageal cancer and amplification of the human cyclin D gene CCND1/PRAD1. Br J Cancer. 71:64–68. 1995.PubMed/NCBI
17	Blagosklonny MV and Pardee AB: The restriction point of the cell cycle. Cell Cycle. 1:103–110. 2002. View Article : Google Scholar : PubMed/NCBI
18	Filmus J, Robles AI, Shi W, et al: Induction of cyclin D1 overexpression by activated ras. Oncogene. 9:3627–3633. 1994.PubMed/NCBI
19	Aktas H, Cai H and Cooper GM: Ras links growth factor signaling to the cell cycle machinery via regulation of cyclin D1 and the cdk inhibitor p27KIP1. Mol Cell Biol. 17:3850–3857. 1997.PubMed/NCBI
20	Winston JT, Coats SR, Wang YZ and Pledger WJ: Regulation of the cell cycle machinery by oncogenic ras. Oncogene. 12:127–134. 1996.PubMed/NCBI
21	Chang HY and Yang X: Proteases for cell suicide: functions and regulation of caspases. Microbiol Mol Biol Rev. 64:821–846. 2000. View Article : Google Scholar : PubMed/NCBI
22	Youle RJ and Strasser A: The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol. 9:47–59. 2008. View Article : Google Scholar : PubMed/NCBI