Skip to main content

Advertisement

SpringerLink
Log in

Tamoxifen Downregulates the Expression of Notch1 and DLL1 Genes in MKN-45 Gastric Cancer Cells

  • Original Research
  • Published:
Journal of Gastrointestinal Cancer Aims and scope Submit manuscript

Abstract

Purpose

Gastric cancer is one of the most prevalent cancers worldwide and the second most common cause for cancer associated mortality. Anti-tumor effects of tamoxifen in breast cancer are well-established. However, no study has so far investigated the effects of tamoxifen on gene expression of Notch1 and DLL1 in gastric cancer cell line. The present study was conducted to explore the effects of tamoxifen, as a repurposed drug, on gene expression of Notch1 and DLL1 in MKN-45, a gastric cancer cell line.

Methods

MKN-45 cells were cultured in DMEM/F12 medium containing 10% FBS. Cytotoxic effects of tamoxifen on these cells at various concentrations were evaluated by trypan blue exclusion assay. For gene expression analysis, the cells were first incubated with 100 μM tamoxifen followed by total RNA extraction from treated and control cells. Then, cDNA was synthesized. Quantitative real-time PCR using specific primers for Notch1 and DLL1 was performed to assess the effect of tamoxifen on the transcript of them.

Results

Treatment with tamoxifen decreased viability of MKN-45 cells in a dose-dependent manner. CC50 was estimated to be around 200 μM. Also, tamoxifen at the dose of 100 μM could significantly downregulate mRNA levels of both Notch1 and DLL1 genes as compared with untreated cells by 24% and 92%, respectively.

Conclusion

Based on these results, tamoxifen interferes with Notch signaling pathway through downregulating the expression of Notch1 and DLL1 genes and this could be regarded as a mechanism for its anti-cancer effects in this malignant disease.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Data Availability

The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions.

Abbreviations

DLL1:

Delta like canonical Notch ligand 1

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

cDNA:

Complementary DNA

PCR:

Polymerase chain reaction

CC50 :

50% cytotoxic concentration

References

  1. Xue H, Li J, Xie H, Wang Y. Review of drug repositioning approaches and resources. Int J Biol Sci. 2018;14(10):1232–44.

    Article  CAS  Google Scholar 

  2. Lee HS, Kim WH, Kwak Y, Koh J, Bae JM, Kim K-M, et al. Molecular testing for gastrointestinal cancer. J Pathol Transl Med. 2017;51(2):103–21.

    Article  Google Scholar 

  3. Li B, Liu H-Y, Guo S-H, Sun P, Gong F-M, Jia B-Q. Detection of microsatellite instability in gastric cancer and dysplasia tissues. Int J Clin Exp Med. 2015;8(11):21442–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Song Z, Wu Y, Yang J, Yang D, Fang X. Progress in the treatment of advanced gastric cancer. Tumor Biol. 2017;39(7):1010428317714626.

    Google Scholar 

  5. Sun Z, Wang Q, Yu X, Ou C, Yao L, Liu K, et al. Risk factors associated with splenic hilar lymph node metastasis in patients with advanced gastric cancer in northwest China. Int J Clin Exp Med. 2015;8(11):21358–64.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Kim J, Yum S, Kang C, Kang S-J. Gene-gene interactions in gastrointestinal cancer susceptibility. Oncotarget. 2016;7(41):67612–25.

    Article  Google Scholar 

  7. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65(2):87–108.

    Article  Google Scholar 

  8. Ho SWT, Tan P. Dissection of gastric cancer heterogeneity for precision oncology. Cancer Sci. 2019;110(11):3405–14.

    Article  CAS  Google Scholar 

  9. Kang W, Zheng X, Wang P, Guo S. Deguelin exerts anticancer activity of human gastric cancer MGC-803 and MKN-45 cells in vitro. Int J Mol Med. 2018;41(6):3157–66.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Wei B, Huang Q, Huang S, Mai W, Zhong X. Trichosanthin-induced autophagy in gastric cancer cell MKN-45 is dependent on reactive oxygen species (ROS) and NF-κB/p53 pathway. J Pharmacol Sci. 2016;131(2):77–83.

    Article  CAS  Google Scholar 

  11. Jiang AG, Yu H, Huang JA. Expression and clinical significance of the phosphatidylinositol 3-kinase/protein kinase B signal transduction pathway in non-small cell lung carcinoma. Oncol Lett. 2014;8(2):601–7.

    Article  Google Scholar 

  12. Zhou W, Fu X, Zhang L, Zhang J, Huang X, Lu X, et al. The AKT1/NF-kappaB/Notch1/PTEN axis has an important role in chemoresistance of gastric cancer cells. Cell Death Dis. 2013;4(10):e847.

    Article  CAS  Google Scholar 

  13. Gelmann EP Tamoxifen for the treatment of malignancies other than breast and endometrial carcinoma. In: Seminars in oncology, 1997. vol 1 Suppl 1. pp S1-65-S61-70

  14. Wei G, Chang Y, Zheng J, He S, Chen N, Wang X, et al. Notch1 silencing inhibits proliferation and invasion in SGC-7901 gastric cancer cells. Mol Med Rep. 2014;9(4):1153–8.

    Article  CAS  Google Scholar 

  15. Piazzi G, Fini L, Selgrad M, Garcia M, Daoud Y, Wex T, et al. Epigenetic regulation of Delta-Like1 controls Notch1 activation in gastric cancer. Oncotarget. 2011;2(12):1291–301.

    Article  Google Scholar 

  16. Bettinsoli P, Ferrari-Toninelli G, Bonini S, Prandelli C, Memo M. Notch ligand Delta-like 1 as a novel molecular target in childhood neuroblastoma. BMC Cancer. 2017;17(1):352.

    Article  CAS  Google Scholar 

  17. Wang X, Chen Q, Huang X, Zou F, Fu Z, Chen Y, et al. Effects of 17β-estradiol and tamoxifen on gastric cancer cell proliferation and apoptosis and ER-α36 expression. Oncol Lett. 2017;13(1):57–62.

    Article  Google Scholar 

  18. Gallo MA, Kaufman D Antagonistic and agonistic effects of tamoxifen: significance in human cancer. In: Seminars in oncology, 1997. vol 1 Suppl 1. pp S1-71-S71-80

  19. Charpentier M, Martin S. Interplay of stem cell characteristics, EMT, and microtentacles in circulating breast tumor cells. Cancers. 2013;5(4):1545–65.

    Article  Google Scholar 

  20. Xiao Y-F, Yong X, Tang B, Qin Y, Zhang J-W, Zhang D, et al. Notch and Wnt signaling pathway in cancer: crucial role and potential therapeutic targets. Int J Oncol. 2016;48(2):437–49.

    Article  CAS  Google Scholar 

  21. Farhangian M, Azarafrouz F, Fallahi H, Akrami H. Investigating the Effect of Ibuprofen on DLL1 and NOTCH1 expression in gastric cancer stem cells derived from MKN-45 cell line. Middle East J Cancer. 2019;10(4):292–8.

    CAS  Google Scholar 

  22. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods. 2001;25(4):402–8.

    Article  CAS  Google Scholar 

  23. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre L, Jemal A. 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 

  24. Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, et al. Cancer statistics in China, 2015. CA Cancer J Clin. 2016;66(2):115–32.

    Article  Google Scholar 

  25. Shan C, Zhang Y, Hao X, Gao J, Chen X, Wang K. Biogenesis, functions and clinical significance of circRNAs in gastric cancer. Mol Cancer. 2019;18(1):136.

    Article  Google Scholar 

  26. Nagini S. Carcinoma of the stomach: a review of epidemiology, pathogenesis, molecular genetics and chemoprevention. World J Gastrointest Oncol. 2012;4(7):156–69.

    Article  Google Scholar 

  27. Yang Z, Guo X, Li G, Shi Y, Li L. Long noncoding RNAs as potential biomarkers in gastric cancer: opportunities and challenges. Cancer Lett. 2016;371(1):62–70.

    Article  CAS  Google Scholar 

  28. Halifu Y, Liang J, Zeng X, Ding Y, Zhang X, Jin T, et al. Wnt1 and SFRP1 as potential prognostic factors and therapeutic targets in cutaneous squamous cell carcinoma. Genet Mol Res. 2016;15(2):8187.

    Article  Google Scholar 

  29. Rizzo P, Miao H, D’Souza G, Osipo C, Yun J, Zhao H, et al. Cross-talk between notch and the estrogen receptor in breast cancer suggests novel therapeutic approaches. Cancer Res. 2008;68(13):5226–35.

    Article  CAS  Google Scholar 

  30. Huang T, Zhou Y, Cheng AS, Yu J, To KF, Kang W. NOTCH receptors in gastric and other gastrointestinal cancers: oncogenes or tumor suppressors? Mol Cancer. 2016;15(1):80.

    Article  Google Scholar 

  31. Pellegrinet L, Rodilla V, Liu Z, Chen S, Koch U, Espinosa L, et al. Dll1-and dll4-mediated notch signaling are required for homeostasis of intestinal stem cells. Gastroenterology. 2011;140(4):1230–1240.e1237.

    Article  CAS  Google Scholar 

  32. Piazzi G, Bazzoli F, Ricciardiello L. Epigenetic silencing of Notch signaling in gastrointestinal cancers. Cell Cycle. 2012;11(23):4323–7.

    Article  CAS  Google Scholar 

  33. Zhu H, Zhou X, Redfield S, He Z, Lewin J, Miele L. Elevated expression of Notch1 is associated with metastasis of human malignancies. Int J Surg Pathol. 2013;21(5):449–54.

    Article  Google Scholar 

  34. Hsu K-W, Hsieh R-H, Huang K-H, Fen-Yau Li A, Chi C-W, Wang T-Y, et al. Activation of the Notch1/STAT3/Twist signaling axis promotes gastric cancer progression. Carcinogenesis. 2012;33(8):1459–67.

    Article  CAS  Google Scholar 

  35. Guo L-Y, Li Y-M, Qiao L, Liu T, Du Y-Y, Zhang J-Q, et al. Notch2 regulates matrix metallopeptidase 9 via PI3K/AKT signaling in human gastric carcinoma cell MKN-45. World J Gastroenterol: WJG. 2012;18(48):7262–70.

    Article  Google Scholar 

Download references

Acknowledgments

The authors are profoundly grateful to Behrouz Moradi, Kiumars Mahdizadeh and Diba Farahani for their contributions in completing this study.

Author information

Authors and Affiliations

  1. Department of Biology, Faculty of Science, Razi University, Kermanshah, Iran

    Faranak Khanipouyani

  2. Gastroenterohepatology Research Center, Shiraz University of Medical Sciences, Shiraz, 71935-1311, Iran

    Hassan Akrami

Authors
  1. Faranak Khanipouyani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hassan Akrami
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Faranak Khanipouyani contributed to experimental works and writing the manuscript, Hassan Akrami designed and directed the investigation and revised the manuscript.

Corresponding author

Correspondence to Hassan Akrami.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethics Approval

Not applicable

Consent to Participate

Not applicable

Consent for Publication

Authors have read and approved the contents of the final manuscript, and all have agreed to publicate this manuscript.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Khanipouyani, F., Akrami, H. Tamoxifen Downregulates the Expression of Notch1 and DLL1 Genes in MKN-45 Gastric Cancer Cells. J Gastrointest Canc 52, 922–927 (2021). https://doi.org/10.1007/s12029-020-00511-y

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12029-020-00511-y

Key words

Search

Navigation