Summary
The purpose of this study was to verify that a combination of mild hyperthermia and docetaxel chemotherapy produces synergistic antitumor effects and to explore the action mechanisms of this treatment approach. The effects of docetaxel on the proliferation of cells from the estrogen receptor (ER)-positive human breast cancer cell line MCF-7 and the ER-negative human breast cancer cell line MDA-MB-453 were examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and effective experimental concentrations of docetaxel were determined. The effects of mild hyperthermia plus docetaxel therapy on apoptosis rate in the MCF-7 and MDA-MB-453 human breast cancer cell lines were analyzed by using flow cytometry with Annexin-V fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. The effects of these combined treatments on cell cycle progression in the MCF-7 and MDA-MB-453 human breast cancer cell lines were examined by using flow cytometry. The effects of these combined treatments on the expression of apoptosis-related proteins and proteins in the mitogen-activated protein kinase (MAPK) pathways were analyzed by using Western blotting. The effects of these combined treatments on the expression of the heat shock protein 70 (HSP70) and the multi-drug resistance (MDR) gene product P-glycoprotein (Pgp) were examined by using Western blotting. The results showed that the half-maximal inhibitory concentration (IC50) of docetaxel for MCF-7 and MDA-MB-453 cells was 19.57±1.12 and 21.64±2.31 μmol/L respectively. Mild hyperthermia with docetaxel therapy could increase apoptosis rate in the MCF-7 and MDA-MB-453 cells. Apoptosis rate in MCF-7 and MDA-MB-453 cells was increased from (23.66±3.59)% and (18.51±3.17)% in docetaxel treatment group to (47.12±6.73)% and (55.16±7.42)% in mild hyperthermia plus docetaxel group, indicating that the mild hyperthermia and docetaxel therapeutic approaches exhibited significant synergistic antitumor effects. Treatments of mild hyperthermia plus docetaxel induced G2/M cell cycle arrest in the MCF-7 and MDA-MB-453 cells. Western blotting demonstrated that proteins in the MAPK pathway were expressed at higher levels in docetaxel-treated cells following mild hypothermia than those in cells treated with docetaxel alone. As compared with blank control group, cells from the mild hyperthermia plus docetaxel group exhibited significantly decreased B-cell lymphoma 2 (Bcl-2) protein expression but slightly increased Bcl-2-associated X protein (Bax) expression. Western blotting results revealed that HSP70 and Pgp expression levels were significantly increased following mild hypothermia. It was concluded that treatments of mild hyperthermia plus docetaxel inhibited the proliferation of human breast cancer cells, promoted apoptosis of breast cancer cells, and produced synergistic antitumor effects.
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References
Vasconcelos A, Medeiros R, Veiga I, et al. Analysis of estrogen receptor polymorphism in codon 325 by PCR-SSCP in breast cancer: association with lymph node metastasis. Breast J, 2002,8(4):226–229
Yamada Y, Itoh Y, Aoki S, et al. Preliminary results of M-VAC chemotherapy combined with mild hyperthermia, a new therapeutic strategy for advanced or metastatic transitional cell carcinoma of the urothelium. Cancer Chemother Pharmacol, 2009,64(6):1079–1083
Mohamed F, Marchettini P, Stuart OA, et al. Thermal enhancement of new chemotherapeutic agents at moderate hyperthermia. Ann Surg Oncol, 2003,10(4):463–468
Klostergaard J, Leroux ME, Auzenne E, et al. Hyperthermia engages the intrinsic apoptotic pathway by enhancing upstream caspase activation to overcome apoptotic resistance in MCF-7 breast adenocarcinoma cells. J Cell Biochem, 2006,98(2):356–369
Peer AJ, Grimm MJ, Zynda ER, et al. Diverse immune mechanisms may contribute to the survival benefit seen in cancer patients receiving hyperthermia. Immunol Res, 2010,46(1–3):137–154
Rong Y, Mack P. Apoptosis induced by hyperthermia in Dunn osteosarcoma cell line in vitro. Int J Hyperthermia, 2000,16(1):19–27
Saga T, Sakahara H, Nakamoto Y, et al. Enhancement of the therapeutic outcome of radio-immunotherapy by combination with whole-body mild hyperthermia. Eur J Cancer, 2001,37(11):1429–1434
Sawaji Y, Sato T, Takeuchi A, et al. Anti-angiogenic action of hyperthermia by suppressing gene expression and production of tumor-derived vascular endothelial growth factor in vivo and in vitro. Br J Cancer, 2002,86(10):1597–1603
Kanaya Y, Doihara H, Shiroma K, et al. Effect of combined therapy with the antiestrogen agent toremifene and local hyperthermia on breast cancer cells implanted in nude mice. Surg Today, 2008,38(10):911–920
Robins HI, Longo W. Whole body hyperthermia: simple complexities. Intensive Care Med, 1999,25(9):898–900
Wendtner C, Abdel-Rahman S, Baumert J, et al. Treatment of primary, recurrent or inadequately resected high-risk soft-tissue sarcomas (STS) of adults: results of a phase II pilot study (RHT-95) of neoadjuvant chemotherapy combined with regional hyperthermia. Eur J Cancer, 2001,37(13):1609–1616
Hildebrandt B, Drager J, Kerner T, et al. Whole-body hyperthermia in the scope of von Ardenne’s systemic cancer multistep therapy (sCMT) combined with chemotherapy in patients with metastatic colorectal cancer: a phase I/II study. Int J Hyperthermia, 2004,20(3): 317–333
Jones EL, Oleson JR, Prosnitz LR, et al. Randomized trial of hyperthermia and radiation for superficial tumors. J Clin Oncol, 2005,23(13):3079–3085
Issels RD, Schlemmer M, Lindner LH. The role of hyperthermia in combined treatment in the management of soft tissue sarcoma. Curr Oncol Rep, 2006,8(4):305–309
Takahashi I, Emi Y, Hasuda S, et al. Clinical application of hyperthermia combined with anticancer drugs for the treatment of solid tumors. Surgery, 2002,131(1 Suppl):S78–S84
Nakano H, Kurihara K, Okamoto M, et al. Heat-induced apoptosis and p53 in cultured mammalian cells. Int J Radiat Biol, 1997,71(5):519–529
Zhu HM, Wang N, Huang X, et al. The effects of hyperthermia on apoptosis in human colonic carcinoma cell line Lovo. Yixue Yanjiusheng Xuebao (Chinese), 2005,18(5):399–401
Zhang L, Liu RG, Zhou JP, et al. Hyperthemia-induced apoptosis of mouse HepA cell line. J Huazhong Univ Sci Tech [Health Sci] (Chinese),2002,31(6):621
Chen WX, Gu ZY, Li YM, et al. Hyperthemia causes apoptosisin human gastric cancer cell line MKN28. Zhongliu Shiyong Zazhi (Chinese), 2001,16(2):126–128
Whyte J, Bergin O, Bianchi A, et al. Key signaling nodes in mammary gland development and cancer. Mitogen-activated protein kinase signaling in experimental models of breast cancer progression and in mammary gland development. Breast Cancer Res, 2009,11(5):209
Bhalla KN. Microtubule-targeted anticancer agents and apoptosis. Oncogene, 2003,22(56):9075–9086
Tabuchi Y, Matsuoka J, Gunduz M, et al. Resistance to paclitaxel therapy is related with Bcl-2 expression through an estrogen receptor mediated pathway in breast cancer. Int J Oncol, 2009,34(2):313–319
Vilaboa NE, Galan A, Troyano A, et al. Regulation of multidrug resistance 1 (MDR1)/P-glycoprotein gene expression and activity by heat-shock transcription factor 1 (HSF1). J Biol Chem, 2000,275(32):24 970–24 976
Kim SH, Yeo GS, Lim YS, et al. Suppression of multidrug resistance via inhibition of heat shock factor by quercetin in MDR cells. Exp Mol Med, 1998,30(2):87–92
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Lv, F., Yu, Y., Zhang, B. et al. Inhibitory effects of mild hyperthermia plus docetaxel therapy on ER(+/−) breast cancer cells and action mechanisms. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 33, 870–876 (2013). https://doi.org/10.1007/s11596-013-1214-8
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DOI: https://doi.org/10.1007/s11596-013-1214-8