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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
  • 28 Downloads

Abstract

Purpose

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).

Methods

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.

Results

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).

Conclusion

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.

Keywords

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

Notes

Acknowledgments

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.

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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

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