Altered Expression of CD44, SIRT1, CXCR4, miR-21, miR-34a, and miR-451 Genes in MKN-45 Cell Line After Docetaxel Treatment

  • Maryam Motamedi
  • Farnaz Razmkhah
  • Leila Rezakhani
  • Sorayya GhasemiEmail author
Original Research



Today it is known that the gene expression profile of cancer stem cells differs from other cancer cells, which may lead to the resistance to routine treatments. The aim of this study was to investigate the effect of docetaxel (DOC) treatment on CD44+ cell frequency in human gastric cancer (GC) MKN-45 cell line and its effect on expression levels of SIRT1, CXCR4, microRNA (miR)-21, miR-451, and miR-34a that are closely correlated with the chemoresistance or self-renewal of cancer stem cells (CSCs).


The cytotoxic effect of DOC on MKN-45 cell line was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)-assay. The frequency of CD44+ cells was measured by flow cytometry in the treated and control groups. The expression level of SIRT1, CXCR4, miR-21, miR-451, and miR-34a was assessed in DOC-treated and non-treated cells using quantitative real-time PCR. Data were analyzed using Statistical Package for the Social Sciences (SPSS) software.


The half-maximal inhibitory concentration (IC50) of DOC was 10 μg/ml after 48 h. Flow cytometry showed a significant increase in CD44+ cells after treatment with DOC (94.3%) when compared with non-treated cells (84.6%) (P < 0.01). The expression of SIRT1, CXCR4, and miR-21 was up-regulated (1.4-fold, 6.7-fold, and 1.22-fold, respectively, P < 0.05) in DOC-treated cells relative to non-treated cells, while miR-451 and miR-34a were down-regulated (0.14-fold and 0.36-fold, respectively, P < 0.05).


DOC treatment affected CD44+ cell frequency in MKN-45 cell line and induced significant changes in the expression of SIRT1, CXCR4, miR-21, miR-451, and miR-34a that are implicated in stemness and chemo-radioresistance, which might offer new insights for future GC therapies.


CD44 SIRT1 CXCR4 miR-21 miR-451 miR-34a Docetaxel Gastric cancer cell line (MKN-45) 



The authors give special thanks to Dr. Akram Alizadeh and all the teachers at the Cellular and Molecular Research Center of Shahrekord University of Medical Sciences for their help and support.

Funding Information

This work was supported by grant no. 2180 from Shahrekord University of Medical Sciences.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. 1.
    Li K, Dan Z, Nie Y-Q. Gastric cancer stem cells in gastric carcinogenesis, progression, prevention and treatment. World J Gastroenterol. 2014;20(18):5420–6.CrossRefGoogle Scholar
  2. 2.
    Wang H, Hajar A, Li S, Chen X, Parissenti AM, Brindley DN, et al. Multiple mechanisms underlying acquired resistance to taxanes in selected docetaxel-resistant MCF-7 breast cancer cells. BMC Cancer. 2014;14(1):37.CrossRefGoogle Scholar
  3. 3.
    Bitarte N, Bandres E, Boni V, Zarate R, Rodriguez J, Gonzalez-Huarriz M, et al. MicroRNA-451 is involved in the self-renewal, tumorigenicity, and chemoresistance of colorectal cancer stem cells. Stem Cells. 2011;29(11):1661–71.CrossRefGoogle Scholar
  4. 4.
    Liu J, Ma L, Xu J, Liu C, Zhang J, Liu J, et al. Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties. Int J Oncol. 2013;42(2):453–9.CrossRefGoogle Scholar
  5. 5.
    Fu Y, Du P, Zhao J, Ce H, Qin Y, Huang G. Gastric cancer stem cells: mechanisms and therapeutic approaches. Yonsei Med J. 2018;59(10):1150–8.CrossRefGoogle Scholar
  6. 6.
    Nishikawa S, Konno M, Hamabe A, Hasegawa S, Kano Y, Ohta K, et al. Aldehyde dehydrogenasehigh gastric cancer stem cells are resistant to chemotherapy. Int J Oncol. 2013;42(4):1437–42.CrossRefGoogle Scholar
  7. 7.
    Tamada M, Nagano O, Tateyama S, Ohmura M, Yae T, Ishimoto T, et al. Modulation of glucose metabolism by CD44 contributes to antioxidant status and drug resistance in cancer cells. Cancer Res. 2012;72(6):1438–48.CrossRefGoogle Scholar
  8. 8.
    Wang W, Dong L-P, Zhang N, Zhao C-H. Role of cancer stem cell marker CD44 in gastric cancer: a meta-analysis. Int J Clin Exp Med. 2014;7(12):5059.Google Scholar
  9. 9.
    Golestaneh AF, Atashi A, Langroudi L, Shafiee A, Ghaemi N, Soleimani M. miRNAs expressed differently in cancer stem cells and cancer cells of human gastric cancer cell line MKN-45. Cell Biochem Funct. 2012;30(5):411–8.CrossRefGoogle Scholar
  10. 10.
    Li X, Yao R, Yue L, Qiu W, Qi W, Liu S, et al. FOXM 1 mediates resistance to docetaxel in gastric cancer via up-regulating Stathmin. J Cell Mol Med. 2014;18(5):811–23.CrossRefGoogle Scholar
  11. 11.
    Qiu X, Wang W, Li B, Cheng B, Lin K, Bai J, et al. Targeting Ezh2 could overcome docetaxel resistance in prostate cancer cells. BMC Cancer. 2019;19(1):27.CrossRefGoogle Scholar
  12. 12.
    Bekaii-Saab T, El-Rayes B. Identifying and targeting cancer stem cells in the treatment of gastric cancer. Cancer. 2017;123(8):1303–12.CrossRefGoogle Scholar
  13. 13.
    Wang Z, Chen W. Emerging roles of SIRT1 in cancer drug resistance. Genes Cancer. 2013;4(3–4):82–90.CrossRefGoogle Scholar
  14. 14.
    Feng Y-H, Tsao C-J. Emerging role of microRNA-21 in cancer. Biomed Rep. 2016;5(4):395–402.CrossRefGoogle Scholar
  15. 15.
    Sekar D, Krishnan R, Panagal M, Sivakumar P, Gopinath V, Basam V. Deciphering the role of microRNA 21 in cancer stem cells (CSCs). Genes Dis. 2016;3(4):277–81.CrossRefGoogle Scholar
  16. 16.
    Wang Z, Wang W, Huang K, Wang Y, Li J, Yang X. MicroRNA-34a inhibits cells proliferation and invasion by downregulating Notch1 in endometrial cancer. Oncotarget. 2017;8(67):111258.Google Scholar
  17. 17.
    Garofalo M, Croce CM. MicroRNAs as therapeutic targets in chemoresistance. Drug Resist Updat. 2013;16(3–5):47–59.CrossRefGoogle Scholar
  18. 18.
    Yokozaki H. Molecular characteristics of eight gastric cancer cell lines established in Japan. Pathol Int. 2000;50(10):767–77.CrossRefGoogle Scholar
  19. 19.
    Nguyen PH, Giraud J, Chambonnier L, Dubus P, Wittkop L, Belleannée G, et al. Characterization of biomarkers of tumorigenic and chemoresistant cancer stem cells in human gastric carcinoma. Clin Cancer Res. 2017;23(6):1586–97.CrossRefGoogle Scholar
  20. 20.
    Dallas NA, Xia L, Fan F, Gray MJ, Gaur P, Van Buren G, et al. Chemoresistant colorectal cancer cells, the cancer stem cell phenotype, and increased sensitivity to insulin-like growth factor-I receptor inhibition. Cancer Res. 2009;69(5):1951–7.CrossRefGoogle Scholar
  21. 21.
    Phillips TM, McBride WH, Pajonk F. The response of CD24−/low/CD44+ breast cancer–initiating cells to radiation. J Natl Cancer Inst. 2006;98(24):1777–85.CrossRefGoogle Scholar
  22. 22.
    Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW, Guba M, et al. Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell. 2007;1(3):313–23.CrossRefGoogle Scholar
  23. 23.
    Dylla SJ, Beviglia L, Park I-K, Chartier C, Raval J, Ngan L, et al. Colorectal cancer stem cells are enriched in xenogeneic tumors following chemotherapy. PLoS One. 2008;3(6):e2428.CrossRefGoogle Scholar
  24. 24.
    Han M, Wang Y, Liu M, Bi X, Bao J, Zeng N, et al. MiR-21 regulates epithelial-mesenchymal transition phenotype and hypoxia-inducible factor-1α expression in third-sphere forming breast cancer stem cell-like cells. Cancer Sci. 2012;103(6):1058–64.CrossRefGoogle Scholar
  25. 25.
    Shi G-h, Ye D-w, X-d Y, S-l Z, Dai B, H-l Z, et al. Involvement of microRNA-21 in mediating chemo-resistance to docetaxel in androgen-independent prostate cancer PC3 cells. Acta Pharmacol Sin. 2010;31(7):867–73.CrossRefGoogle Scholar
  26. 26.
    Choi G, Lee J, Ji JY, Woo J, Kang NS, Cho SY, et al. Discovery of a potent small molecule SIRT1/2 inhibitor with anticancer effects. Int J Oncol. 2013;43(4):1205–11.CrossRefGoogle Scholar
  27. 27.
    Chen X, Sun K, Jiao S, Cai N, Zhao X, Zou H, et al. High levels of SIRT1 expression enhance tumorigenesis and associate with a poor prognosis of colorectal carcinoma patients. Sci Rep. 2014;4:7481.CrossRefGoogle Scholar
  28. 28.
    Zhu H, Xia L, Zhang Y, Wang H, Xu W, Hu H, et al. Activating transcription factor 4 confers a multidrug resistance phenotype to gastric cancer cells through transactivation of SIRT1 expression. PLoS One. 2012;7(2):e31431.CrossRefGoogle Scholar
  29. 29.
    Zhao B-C, Wang Z-J, Mao W-Z, Ma H-C, Han J-G, Zhao B, et al. CXCR4/SDF-1 axis is involved in lymph node metastasis of gastric carcinoma. World J Gastroenterol. 2011;17(19):2389.CrossRefGoogle Scholar
  30. 30.
    Dalerba P, Clarke MF. Cancer stem cells and tumor metastasis: first steps into uncharted territory. Cell Stem Cell. 2007;1(3):241–2.CrossRefGoogle Scholar
  31. 31.
    Gatti M, Pattarozzi A, Bajetto A, Würth R, Daga A, Fiaschi P, et al. Inhibition of CXCL12/CXCR4 autocrine/paracrine loop reduces viability of human glioblastoma stem-like cells affecting self-renewal activity. Toxicology. 2013;314(2–3):209–20.CrossRefGoogle Scholar
  32. 32.
    Xie L, Wei J, Qian X, Chen G, Yu L, Ding Y, et al. CXCR4, a potential predictive marker for docetaxel sensitivity in gastric cancer. Anticancer Res. 2010;30(6):2209–16.Google Scholar
  33. 33.
    Liu C, Tang DG. MicroRNA regulation of cancer stem cells. Cancer Res. 2011;71(18):5950–4.CrossRefGoogle Scholar
  34. 34.
    Bandres E, Bitarte N, Arias F, Agorreta J, Fortes P, Agirre X, et al. microRNA-451 regulates macrophage migration inhibitory factor production and proliferation of gastrointestinal cancer cells. Clin Cancer Res. 2009;15(7):2281–90.CrossRefGoogle Scholar
  35. 35.
    Gal H, Pandi G, Kanner AA, Ram Z, Lithwick-Yanai G, Amariglio N, et al. MIR-451 and imatinib mesylate inhibit tumor growth of glioblastoma stem cells. Biochem Biophys Res Commun. 2008;376(1):86–90.CrossRefGoogle Scholar
  36. 36.
    Bu P, Chen K-Y, Chen JH, Wang L, Walters J, Shin YJ, et al. A microRNA miR-34a-regulated bimodal switch targets Notch in colon cancer stem cells. Cell Stem Cell. 2013;12(5):602–15.CrossRefGoogle Scholar
  37. 37.
    Siemens H, Jackstadt R, Kaller M, Hermeking H. Repression of c-Kit by p53 is mediated by miR-34 and is associated with reduced chemoresistance, migration and stemness. Oncotarget. 2013;4(9):1399.CrossRefGoogle Scholar
  38. 38.
    Li Y, Guessous F, Zhang Y, DiPierro C, Kefas B, Johnson E, et al. MicroRNA-34a inhibits glioblastoma growth by targeting multiple oncogenes. Cancer Res. 2009;69(19):7569–76.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Cellular and Molecular Research Center, Basic Health Sciences InstituteShahrekord University of Medical SciencesShahrekordIran
  2. 2.Hematology Research CenterShiraz University of Medical SciencesShirazIran
  3. 3.Department of Tissue Engineering, School of Advanced TechnologiesShahrekord University of Medical SciencesShahrekordIran

Personalised recommendations