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Silymarin inhibited DU145 cells by activating SLIT2 protein and suppressing expression of CXCR4

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Abstract

Among other cancers, prostate cancer is globally the second most rampant one with the incidence of 29.4% among men. SLIT2/ROBO1 signaling is very crucial pathway causally implicated in many cancers and reported to inhibit a variety of cancer cell types. CXCR4 is a chemokine receptor implicated in cancer progression. Silymarin is a phytochemical, of which anti-carcinogenic activity was suggested in various cancers, including prostate cancer. However, there are no studies examining the effect of silymarin on SLIT2–Robo1–CXCR4 axis. Herein, our goal is to explore cytotoxic and morphological effects of silymarin on DU145 cells and to reveal its role in Slit2/Robo and CXCR1 pathway. First, 24, 48 and 72 h-long cytotoxicity tests were performed for dose analysis of silymarin, followed H–E stain for morphological evaluation with varying doses of silymarin. Afterward, western blot and immunocytochemistry analyses were carried out for SLIT2, ROBO1 and CXCR4 proteins. According to MTT analysis, IC50 concentrations for silymarin were 315, 126 and 70 µM against DU145 cells for 24, 48 and 72 h treatments. In H–E, several apoptotic hallmarks, including, condensed, kidney-shaped and eccentric nuclei, membrane blebbings and apoptotic body formations were observed. Silymarin increased the expressions of SLIT2 and ROBO1 while decreased CXCR4 when compared to control group in immunocytochemistry and Western blot. To summarize, silymarin inhibited DU145 cells dose-dependently by activating SLIT2 protein and inhibiting expression of CXCR4. This study is the first examining the interplay between Slit2–Robo1–CXCR4 proteins and silymarin in DU145 cells. We believe that our study will provide new insights for future studies.

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Data availability

All data generated or analyzed during the present study are included in this published article.

References

  1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68(6):394–424.

    Article  Google Scholar 

  2. Gara RK, Kumari S, Ganju A, Yallapu MM, Jaggi M, Chauhan SC. Slit/Robo pathway: a promising therapeutic target for cancer. Drug Discov Today. 2015;20(1):156–64.

    Article  CAS  Google Scholar 

  3. Andrews W, Liapi A, Plachez C, Camurri L, Zhang J, Mori S, et al. Robo1 regulates the development of major axon tracts and interneuron migration in the forebrain. Development. 2006;133(11):2243–52.

    Article  CAS  Google Scholar 

  4. Shtivelman E, Beer TM, Evans CP. Molecular pathways and targets in prostate cancer. Oncotarget. 2014;5(17):7217–59.

    Article  Google Scholar 

  5. Yu J, Cao Q, Yu J, Wu L, Dallol A, Li J, et al. The neuronal repellent SLIT2 is a target for repression by EZH2 in prostate cancer. Oncogene. 2010;29(39):5370–80.

    Article  CAS  Google Scholar 

  6. Teicher BA, Fricker SP. CXCL12 (SDF-1)/CXCR4 pathway in cancer. Clin Cancer Res. 2010;16(11):2927–31.

    Article  CAS  Google Scholar 

  7. Stone KR, Mickey DD, Wunderli H, Mickey GH, Paulson DF. Isolation of a human prostate carcinoma cell line (DU 145). Int J Cancer. 1978;21(3):274–81.

    Article  CAS  Google Scholar 

  8. Ramasamy K, Agarwal R. Multitargeted therapy of cancer by silymarin. Cancer Lett. 2008;269(2):352–62.

    Article  CAS  Google Scholar 

  9. Surai PF. Silymarin as a natural antioxidant: an overview of the current evidence and perspectives. Antioxidants. 2015;4(1):204–47.

    Article  CAS  Google Scholar 

  10. Zi XL, Grasso AW, Kung HJ, Agarwal R. A flavonoid antioxidant, silymarin, inhibits activation of erbB1 signaling and induces cyclin-dependent kinase inhibitors, G1 arrest, and anticarcinogenic effects in human prostate carcinoma DU145 cells. Cancer Res. 1998;58(9):1920–9.

    CAS  PubMed  Google Scholar 

  11. Bhatia N, Agarwal R. Detrimental effect of cancer preventive phytochemicals silymarin, genistein and epigallocatechin 3-gallate on epigenetic events in human prostate carcinoma DU145 cells. Prostate. 2001;46(2):98–107.

    Article  CAS  Google Scholar 

  12. Deep G, Agarwal R. Chemopreventive efficacy of silymarin in skin and prostate cancer. Integr Cancer Ther. 2007;6(2):130–45.

    Article  CAS  Google Scholar 

  13. Dhanalakshmi S, Singh RP, Agarwal C, Agarwal R. Silibinin inhibits constitutive and TNFalpha-induced activation of NF-kappaB and sensitizes human prostate carcinoma DU145 cells to TNFalpha-induced apoptosis. Oncogene. 2002;21(11):1759–67.

    Article  CAS  Google Scholar 

  14. Jiang C, Agarwal R, Lu J. Anti-angiogenic potential of a cancer chemopreventive flavonoid antioxidant, silymarin: inhibition of key attributes of vascular endothelial cells and angiogenic cytokine secretion by cancer epithelial cells. Biochem Biophys Res Commun. 2000;276(1):371–8.

    Article  CAS  Google Scholar 

  15. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1–2):55–63.

    Article  CAS  Google Scholar 

  16. Feng N, Luo J, Guo X. Silybin suppresses cell proliferation and induces apoptosis of multiple myeloma cells via the PI3K/Akt/mTOR signaling pathway. Mol Med Rep. 2016;13(4):3243–8.

    Article  CAS  Google Scholar 

  17. Zheng DD, Wang YC, Zhang DR, Liu ZP, Duan CX, Jia LJ, et al. In vitro antitumor activity of silybin nanosuspension in PC-3 cells. Cancer Lett. 2011;307(2):158–64.

    Article  CAS  Google Scholar 

  18. Mahmoodi N, Motamed N, Paylakhi SH. The Comparison of the effects of silybin and silybin-phosphatidylcholine on viability and ESR expression in human breast cancer T47D cell line. Cell J. 2014;16(3):299–308.

    PubMed  PubMed Central  Google Scholar 

  19. Mahmoodi N, Motamed N, Paylakhi SH, Mahmoodi NO. Comparing the effect of silybin and silybin advanced (TM) on viability and HER2 expression on the human breast cancer SKBR3 cell line by no serum starvation. Iran J Pharm Res. 2015;14(2):521–30.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Zhao Y, Zhou F-L, Li W-P, Wang J, Wang L-J. Slit2-Robo1 signaling promotes the adhesion, invasion and migration of tongue carcinoma cells via upregulating matrix metalloproteinases 2 and 9, and downregulating E-cadherin. Mol Med Rep. 2016;14(3):1901–6.

    Article  CAS  Google Scholar 

  21. Wang B, Xiao Y, Ding BB, Zhang N, Yuan X, Gui L, et al. Induction of tumor angiogenesis by Slit–Robo signaling and inhibition of cancer growth by blocking Robo activity. Cancer Cell. 2003;4(1):19–29.

    Article  Google Scholar 

  22. Tseng RC, Lee SH, Hsu HS, Chen BH, Tsai WC, Tzao C, et al. SLIT2 attenuation during lung cancer progression deregulates beta-catenin and E-cadherin and associates with poor prognosis. Cancer Res. 2010;70(2):543–51.

    Article  CAS  Google Scholar 

  23. Dallol A, Krex D, Hesson L, Eng C, Maher ER, Latif F. Frequent epigenetic inactivation of the SLIT2 gene in gliomas. Oncogene. 2003;22(29):4611–6.

    Article  CAS  Google Scholar 

  24. Dallol A, Morton D, Maher ER, Latif F. SLIT2 axon guidance molecule is frequently inactivated in colorectal cancer and suppresses growth of colorectal carcinoma cells. Cancer Res. 2003;63(5):1054–8.

    CAS  PubMed  Google Scholar 

  25. Dallol A, Da Silva NF, Viacava P, Minna JD, Bieche I, Maher ER, et al. SLIT2, a human homologue of the Drosophila Slit2 gene, has tumor suppressor activity and is frequently inactivated in lung and breast cancers. Cancer Res. 2002;62(20):5874–80.

    CAS  PubMed  Google Scholar 

  26. Latil A, Chene L, Cochant-Priollet B, Mangin P, Fournier G, Berthon P, et al. Quantification of expression of netrins, slits and their receptors in human prostate tumors. Int J Cancer. 2003;103(3):306–15.

    Article  CAS  Google Scholar 

  27. Chatterjee S, Behnam Azad B, Nimmagadda S. The intricate role of CXCR4 in cancer. Adv Cancer Res. 2014;124:31–82.

    Article  CAS  Google Scholar 

  28. Prasad A, Qamri Z, Wu J, Ganju RK. Slit-2/Robo-1 modulates the CXCL12/CXCR4-induced chemotaxis of T cells. J Leukoc Biol. 2007;82(3):465–76.

    Article  CAS  Google Scholar 

  29. Prasad A, Fernandis AZ, Rao Y, Ganju RK. Slit protein-mediated inhibition of CXCR4-induced chemotactic and chemoinvasive signaling pathways in breast cancer cells. J Biol Chem. 2004;279(10):9115–244.

    Article  CAS  Google Scholar 

  30. Marlow R, Strickland P, Lee JS, Wu X, Pebenito M, Binnewies M, et al. SLITs suppress tumor growth in vivo by silencing Sdf1/Cxcr4 within breast epithelium. Cancer Res. 2008;68(19):7819–27.

    Article  CAS  Google Scholar 

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The authors declare that they have no financial competing interests.

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Authors and Affiliations

  1. Department of Histology and Embryology, Faculty of Medicine, Eskisehir Osmangazi University, Eskisehir, Turkey

    Sedat Kacar, Nuriye Ezgi Bektur Aykanat & Varol Sahinturk

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  1. Sedat Kacar
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  2. Nuriye Ezgi Bektur Aykanat
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  3. Varol Sahinturk
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Contributions

SK, EB, VS conceived and designed the study. SK wrote the paper. All authors reviewed and edited the manuscript.

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Correspondence to Sedat Kacar.

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Kacar, S., Bektur Aykanat, N.E. & Sahinturk, V. Silymarin inhibited DU145 cells by activating SLIT2 protein and suppressing expression of CXCR4. Med Oncol 37, 18 (2020). https://doi.org/10.1007/s12032-020-1343-4

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  • DOI: https://doi.org/10.1007/s12032-020-1343-4

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