Effect of RAD51C expression on the chemosensitivity of Eμ-Myc p19Arf‑/‑ cells and its clinical significance in breast cancer

Liao,Shao‑Guang; Liu,Lu; Wang,Ya‑Jie

doi:10.3892/ol.2018.8133

Effect of RAD51C expression on the chemosensitivity of Eμ-Myc p19Arf‑/‑ cells and its clinical significance in breast cancer

Authors:
- Shao‑Guang Liao
- Lu Liu
- Ya‑Jie Wang
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Affiliations: Department of Radiation Oncology, Fuzhou General Hospital, Fuzhou, Fujian 350001, P.R. China, Department of Oncology, Shenyang General Hospital, Shenyang, Liaoning 110000, P.R. China, Department of Oncology, Changhai Hospital, Second Military Medical University, Shanghai 200433, P.R. China
Published online on: February 28, 2018 https://doi.org/10.3892/ol.2018.8133
Pages: 6107-6114
Copyright: © Liao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the chemosensitivity to anti‑cancer drugs of RAD51 paralog C (RAD51C)-deficient Eμ-Myc p19Arf‑/‑ cells, to detect the expression of RAD51C in breast cancer tissues by immunohistochemistry (IHC), and to explore their association with clinicopathological factors. Eμ‑Myc p19Arf‑/‑ cells were stably transfected with retroviruses co‑expressing short hairpin‑RNA against RAD51C and green fluorescent protein (GFP). A single‑cell flow cytometry‑based GFP competition assay was used to assess the change in sensitivity to anti‑cancer drugs. GFP‑negative cells in the same population served as an internal control. In total, tissue samples from 213 cases of breast cancer and 99 adjacent non‑cancerous tissue samples were collected to construct tissue microarrays. IHC was used to detect the expression of RAD51C protein. Relevant clinical information was collected for a correlation analysis. Transfection of RAD51C‑shRNA was demonstrated to effectively reduce the RAD51C protein expression in the Eμ‑Myc p19Arf‑/‑ cells. The sensitivities of the cells to three drugs, camptothecin, cisplatin and olaparib, significantly increased following RAD51C gene knockdown. In breast cancer tissue, RAD51C expression was significantly higher in the Erb‑B2 receptor tyrosine kinase 2 overexpression group. The overall survival time of the patients with RAD51C‑negative expression was longer than that of patients with RAD51C‑positive expression. RAD51C expression was an independent prognostic factor for survival of breast cancer patients. In summary, the results indicate that silencing of RAD51C may represent a potential therapeutic strategy for malignant tumors, and that measuring RAD51C expression by IHC may have prognostic value for breast cancer patients.

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1	Khanna KK and Jackson SP: DNA double-strand breaks: Signaling, repair and the cancer connection. Nat Genet. 27:247–254. 2001. View Article : Google Scholar : PubMed/NCBI
2	Ciccia A and Elledge SJ: The DNA damage response: Making it safe to play with knives. Mol Cell. 40:179–204. 2010. View Article : Google Scholar : PubMed/NCBI
3	Chun J, Buechelmaier ES and Powell SN: Rad51 paralog complexes BCDX2 and CX3 act at different stages in the BRCA1-BRCA2-dependent homologous recombination pathway. Mol Cell Biol. 33:387–395. 2013. View Article : Google Scholar : PubMed/NCBI
4	Badie S, Liao C, Thanasoula M, Barber P, Hill MA and Tarsounas M: RAD51C facilitates checkpoint signaling by promoting CHK2 phosphorylation. J Cell Biol. 185:587–600. 2009. View Article : Google Scholar : PubMed/NCBI
5	Liu Y, Masson JY, Shah R, O'Regan P and West SC: RAD51C is required for Holliday junction processing in mammalian cells. Science. 303:243–246. 2004. View Article : Google Scholar : PubMed/NCBI
6	Kuznetsov S, Pellegrini M, Shuda K, Fernandez-Capetillo O, Liu Y, Martin BK, Burkett S, Southon E, Pati D, Tessarollo L, et al: RAD51C deficiency in mice results in early prophase I arrest in males and sister chromatid separation at metaphase II in females. J Cell Biol. 176:581–592. 2007. View Article : Google Scholar : PubMed/NCBI
7	Meindl A, Hellebrand H, Wiek C, Erven V, Wappenschmidt B, Niederacher D, Freund M, Lichtner P, Hartmann L, Schaal H, et al: Germline mutations in breast and ovarian cancer pedigrees establish RAD51C as a human cancer susceptibilitygene. Nat Genet. 42:410–414. 2010. View Article : Google Scholar : PubMed/NCBI
8	Pelttari LM, Heikkinen T, Thompson D, Kallioniemi A, Schleutker J, Holli K, Blomqvist C, Aittomäki K, Bützow R and Nevanlinna H: RAD51C is a susceptibility gene for ovarian cancer. Hum Mol Genet. 20:3278–3288. 2011. View Article : Google Scholar : PubMed/NCBI
9	Loveday C, Turnbull C, Ruark E, Xicola RM, Ramsay E, Hughes D, Warren-Perry M and Snape K: Breast Cancer Susceptibility Collaboration (UK), Eccles D, et al: Germline RAD51C mutations confer susceptibility to ovarian cancer. Nat Genet. 44:475–476. 2012. View Article : Google Scholar : PubMed/NCBI
10	Vaz F, Hanenberg H, Schuster B, Barker K, Wiek C, Erven V, Neveling K, Endt D, Kesterton I, Autore F, et al: Mutation of the RAD51C gene in a Fanconi anemia-like disorder. Nat Genet. 42:406–409. 2010. View Article : Google Scholar : PubMed/NCBI
11	Forbes SA, Beare D, Gunasekaran P, Leung K, Bindal N, Boutselakis H, Ding M, Bamford S, Cole C, Ward S, et al: COSMIC: Exploring the world's knowledge of somatic mutations in human cancer. Nucleic Acids Res. 3:D805–D811. 2015. View Article : Google Scholar
12	Jiang H, Pritchard JR, Williams RT, Lauffenburger DA and Hemann MT: A mammalian functional-genetic approach to characterizing cancer therapeutics. Nat Chem Biol. 7:92–100. 2011. View Article : Google Scholar : PubMed/NCBI
13	Dickins RA, Hemann MT, Zilfou JT, Simpson DR, Ibarra I, Hannon GJ and Lowe SW: Probing tumor phenotypes using stable and regulated synthetic microRNA precursors. Nat Genet. 37:1289–1295. 2005. View Article : Google Scholar : PubMed/NCBI
14	Jiang H, Reinhardt HC, Bartkova J, Tommiska J, Blomqvist C, Nevanlinna H, Bartek J, Yaffe MB and Hemann MT: The combined status of ATM and p53 link tumor development with therapeutic response. Genes Dev. 23:1895–1909. 2009. View Article : Google Scholar : PubMed/NCBI
15	Pfaffl MW: A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29:e452001. View Article : Google Scholar : PubMed/NCBI
16	Cai MY, Tong ZT, Zheng F, Liao YJ, Wang Y, Rao HL, Chen YC, Wu QL, Liu YH, Guan XY, et al: EZH2 protein: A promising immunomarker for the detection of hepatocellular carcinomas in liver needle biopsies. Gut. 60:967–976. 2011. View Article : Google Scholar : PubMed/NCBI
17	Thacker J: The RAD51 gene family, genetic instability and cancer. Cancer Lett. 219:125–135. 2005. View Article : Google Scholar : PubMed/NCBI
18	Masson JY, Tarsounas MC, Stasiak AZ, Stasiak A, Shah R, McIlwraith MJ, Benson FE and West SC: Identification and purification of two distinct complexes containing the five RAD51 paralogs. Genes Dev. 15:3296–3307. 2001. View Article : Google Scholar : PubMed/NCBI
19	Suwaki N, Klare K and Tarsounas M: RAD51 paralogs: Roles in DNA damage signalling, recombinational repair and tumorigenesis. Semin Cell Dev Biol. 22:898–905. 2011. View Article : Google Scholar : PubMed/NCBI
20	Sinha BK: Topoisomerase inhibitors. A review of their therapeutic potential in cancer. Drugs. 49:11–19. 1995. View Article : Google Scholar : PubMed/NCBI
21	Pommier Y: Topoisomerase I inhibitors: Camptothecins and beyond. Nat Rev Cancer. 6:789–802. 2006. View Article : Google Scholar : PubMed/NCBI
22	Altaha R, Liang X, Yu JJ and Reed E: Excision repair cross complementing-group 1: Gene expression and platinum resistance. Int J Mol Med. 14:959–970. 2004.PubMed/NCBI
23	Boulton S, Kyle S and Durkacz BW: Interactive effects of inhibitors of poly (ADP-ribose. polymerase and DNA-dependent protein kinase on cellular responses to DNA damage. Carcinogenesis. 20:199–203. 1999. View Article : Google Scholar : PubMed/NCBI
24	Haber JE: DNA recombination: The replication connection. Trends Biochem Sci. 24:271–275. 1999. View Article : Google Scholar : PubMed/NCBI
25	Arnaudeau C, Lundin C and Helleday T: DNA double-strand breaks associated with replication forks are predominantly repaired by homologous recombination involving an exchange mechanism in mammalian cells. J Mol Biol. 307:1235–1245. 2001. View Article : Google Scholar : PubMed/NCBI
26	Fukutomi T and Akashi-Tanaka S: Prognostic and predictive factors in the adjuvant treatment of breast cancer. Breast Cancer. 9:95–99. 2002. View Article : Google Scholar : PubMed/NCBI