Expression of Nestin and its relationship with tamoxifen curative effect and prognosis of patients with breast cancer

doi:10.3760/cma.j.issn.1673-422X.2019.04.003

Abstract

Abstract: Objective To investigate the expression of Nestin and its relationship with tamoxifen curative effect and prognosis of patients with breast cancer. Methods A total of 82 patients with breast cancer who received tamoxifen therapy after radical mastectomy in Department of Breast Surgery of Enshi Tujia Miao Autonomous Prefecture Central Hospital of Hubei Province from March 2009 to March 2013 were collected. And the paired cancer tissues and adjacent normal tissues preserved in liquid nitrogen were also collected. Fluorescent quantitative real-time PCR (RT-PCR) and immunohistochemistry were used to detect the content of Nestin mRNA and the expression of Nestin in breast cancer tissues. The expression of Nestin in breast cancer tissues and its relationship with clinicopathologic features were analyzed. Breast cancer cell line MCF-7 was selected, small interfering RNA (siRNA) was used to silence Nestin in MCF-7 cells, and the influence on tamoxifen sensitivity was observed. The relationship of Nestin and epithelial-mesenchymal transition (EMT) was detected using Western blotting. Results The results of RT-PCR indicated that the mRNA level of Nestin in breast cancer tissues was 3.87 times as high as that in paracancerous tissues (6.34±1.56 vs. 1.64±0.52, t=26.140, P＜0.001). Immunohistochemical staining suggested that the positive rate of Nestin in breast cancer tissues was 75.61% (62/82), which was significantly higher than that in paracancerous tissues ［24.39% (20/82)］, with a significant difference (χ2=43.024, P＜0.001). The expression of Nestin was related to lymphatic vessel infiltration (χ2=7.499, P=0.006) and lymph node metastasis (χ2=6.770, P=0.034), and it was not related to the age of patients (χ2=3.242, P=0.072), tumor size (χ2=2.358, P=0.308), histological grade (χ2=0.294, P=0.863), ductal infiltrating status (χ2=0.180, P=0.671) and triple negative breast cancer (χ2=0.142, P=0.706). The analysis of Cox risk ratio model showed that Nestin expression (HR=1.982, P<0.001; HR=1.537, P<0.001), lymphatic vessel invasion (HR=2.502, P<0.001; HR=1.715, P<0.001) and lymph node metastasis (HR=1.818, P<0.001; HR=1.446, P<0.001) were independent prognostic risk factors for progress-free survival and overall survival in breast cancer patients. After Nestin knockout in breast cancer cell line MCF-7, the IC50 value of MCF-7 to tamoxifen decreased from (48.05±2.22) μmol/L to (35.59±2.92) μmol/L, with a significant difference (t=6.489, P=0.003). Silencing Nestin significantly up-regulated E-cadherin (7.21±1.15 vs. 1.02±0.01, t=6.654, P=0.024) and down-regulated the mesenchyme index Vimentin (0.17±0.08 vs. 1.01±0.02, t=25.015, P＜0.001) in MCF-7 cells. Conclusion Nestin is over-expressed in breast cancer, which is associated with reduced efficacy of tamoxifen. Nestin may be a new potential prognostic biomarker for breast cancer.

Key words: Breast neoplasms, Tamoxifen, Nestin, Biomarkers, tumor

Li Yinmou, Li Jing, Peng Shijun. Expression of Nestin and its relationship with tamoxifen curative effect and prognosis of patients with breast cancer[J]. Journal of International Oncology, 2019, 46(4): 205-210.

References

［1］ DeSantis C, Ma J, Bryan L, et al. Breast cancer statistics, 2013［J］. CA Cancer J Clin, 2014, 64(1): 52-62. DOI: 10.3322/caac.21203. ［2］ DeSantis CE, Fedewa SA, Goding Sauer A, et al. Breast cancer statistics, 2015: convergence of incidence rates between black and white women［J］. CA Cancer J Clin, 2016, 66(1): 31-42. DOI: 10.3322/caac.21320. ［3］ Rea D, Handley K, Bowden SJ, et al. aTTom: long-term effects of continuing adjuvant tamoxifen to 10 years versus stopping at 5 years in 6,953 women with early breast cancer［J］. J Clin Oncol, 2013, 31 Suppl 18: 5. ［4］ Catalano S, Giordano C, Panza S, et al. Tamoxifen through GPER upregulates aromatase expression: a novel mechanism sustaining tamoxifenresistant breast cancer cell growth［J］. Breast Cancer Res Treat, 2014, 146(2): 273-285. DOI: 10.1007/s10549-014-3017-4. ［5］ Mc Ilroy M, Fleming FJ, Buggy Y, et al. Tamoxifen-induced ER-alpha-SRC-3 interaction in HER2 positive human breast cancer; a possible mechanism for ER isoform specific recurrence［J］. Endocr Relat Cancer, 2006, 13(4): 1135-1145. DOI: 10.1677/erc.1.01222. ［6］ Piva M, Domenici G, Iriondo O, et al. Sox2 promotes tamoxifen resistance in breast cancer cells［J］. EMBO Mol Med, 2014, 6(1): 66-79. DOI: 10.1002/emmm.201303411. ［7］ Messam CA, Hou J, Major EO. Coexpression of nestin in neural and glial cells in the developing human CNS defined by a human-specific anti-nestin antibody［J］. Exp Neurol, 2000, 161(2): 585-596. DOI: 10.1006/exnr.1999.7319. ［8］ Ishiwata T, Matsuda Y, Naito Z. Nestin in gastrointestinal and other cancers: effects on cells and tumor angiogenesis［J］. World J Gastroenterol, 2011, 17(4): 409-418. DOI: 10.3748/wjg.v17.i4.409. ［9］ Piras F, Ionta MT, Lai S, et al. Nestin expression associates with poor prognosis and triple negative phenotype in locally advanced (T4) breast cancer［J］. Eur J Histochem, 2011, 55(4): e39. DOI: 10.4081/ejh.2011.e39. ［10］ Chou YH, Khuon S, Herrmann H, et al. Nestin promotes the phosphorylationdependent disassembly of vimentin intermediate filaments during mitosis［J］. Mol Biol Cell, 2003, 14(4): 1468-1478. DOI: 10.1091/mbc.e02-08-0545. ［11］ Polyak K. Heterogeneity in breast cancer［J］. J Clin Invest, 2011, 121(10): 3786-3788. DOI: 10.1172/JCI60534. ［12］ Kurokawa H, Lenferink AE, Simpson JF, et al. Inhibition of HER2/neu (erbB-2) and mitogenactivated protein kinases enhances tamoxifen action against HER2-overexpressing, tamoxifen-resistant breast cancer cells［J］. Cancer Res, 2000, 60(20): 5887-5894. ［13］ Joensuu H, KellokumpuLehtinen PL, Bono P, et al. Adjuvant docetaxel or vinorelbine with or without trastuzumab for breast cancer［J］. N Engl J Med, 2006, 354(8): 809-820. DOI: 10.1056/NEJMoa053028. ［14］ Bauer KR, Brown M, Cress RD, et al. Descriptive analysis of estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, and HER2-negative invasive breast cancer, the so-called triple-negative phenotype: a populationbased study from the California cancer registry［J］. Cancer, 2007, 109(9): 1721-1728. DOI: 10.1002/cncr.22618. ［15］ Michalczyk K, Ziman M. Nestin structure and predicted function in cellular cytoskeletal organisation［J］. Histol Histopathol, 2005, 20(2): 665-671. DOI: 10.14670/HH-20.665. ［16］ Dahlstrand J, Lardelli M, Lendahl U. Nestin mRNA expression correlates with the central nervous system progenitor cell state in many, but not all, regions of developing central nervous system［J］. Brain Res Dev Brain Res, 1995, 84(1): 109-129. ［17］ Ehrmann J, Kolár Z, Mokry J. Nestin as a diagnostic and prognostic marker: immunohistochemical analysis of its expression in different tumours［J］. J Clin Pathol, 2005, 58(2): 222-223. DOI: 10.1136/jcp.2004.021238. ［18］ R?gelsperger O, Ekmekcioglu C, J?ger W, et al. Coexpression of the melatonin receptor 1 and nestin in human breast cancer specimens［J］. J Pineal Res, 2009, 46(4): 422-432. DOI: 10.1111/j.1600-079X.2009.00679.x. ［19］ Li H, Cherukuri P, Li N, et al. Nestin is expressed in the basal/myoepithelial layer of the mammary gland and is a selective marker of basal epithelial breast tumors［J］. Cancer Res, 2014, 67(2): 501-510. DOI: 10.1158/00085472.CAN-05-4571. ［20］ Zheng H, Ying H, Yan H, et al. p53 and Pten control neural and glioma stem/progenitor cell renewal and differentiation［J］. Nature, 2011, 455(7216): 1129-1133. DOI: 10.1038/nature07443. ［21］ Zhong B, Wang T, Zou J, et al. Association of the intermediate filament nestin with cancer stage: a meta-analysis based on 223 positive/high nestin cases and 460 negative/low case-free controls［J］. Oncotarget, 2015, 6(26): 22970-22977. DOI: 10.18632/oncotarget.4042. ［22］ Neradil J, Veselska R. Nestin as a marker of cancer stem cells［J］. Cancer Sci, 2015, 106(7): 803-811. DOI: 10.1111/cas.12691. ［23］ Osman WM, Shash LS, Ahmed NS. Emerging role of nestin as an angiogenesis and cancer stem cell marker in epithelial ovarian cancer: immunohistochemical study［J］. Appl Immunohistochem Mol Morphol, 2017, 25(8): 571-580. DOI: 10.1097/PAI.0000000000000338. ［24］ Feng W, Liu S, Zhu R, et al. SOX10 induced Nestin expression regulates cancer stem cell properties of TNBC cells［J］. Biochem Biophys Res Commun, 2017, 485(2): 522-528. DOI: 10.1016/j.bbrc.2017.02.014. ［25］ Chaffer CL, San Juan BP, Lim E, et al. EMT, cell plasticity and metastasis［J］. Cancer Metastasis Rev, 2016, 35(4): 645-654. DOI: 10.1007/s10555-016-9648-7. ［26］ Zhao Z, Lu P, Zhang H, et al. Nestin positively regulates the Wnt/β-catenin pathway and the proliferation, survival and invasiveness of breast cancer stem cells［J］. Breast Cancer Res, 2014, 16(4): 408. DOI: 10.1186/s13058-014-0408-8.

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