Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Dec 29;11(1):97.
doi: 10.3390/cells11010097.

DSTYK Enhances Chemoresistance in Triple-Negative Breast Cancer Cells

Stella C Ogbu  1 , Samuel Rojas  1 , John Weaver  1 , Phillip R Musich  1 , Jinyu Zhang  2 , Zhi Q Yao  2 , Yong Jiang  1
Affiliations

DSTYK Enhances Chemoresistance in Triple-Negative Breast Cancer Cells

Stella C Ogbu et al. Cells. .

Abstract

Breast cancer, as the most prevalent cancer in women, is responsible for more than 15% of new cancer cases and about 6.9% of all cancer-related death in the US. A major cause of therapeutic failure in breast cancer is the development of resistance to chemotherapy, especially for triple-negative breast cancer (TNBC). Therefore, how to overcome chemoresistance is the major challenge to improve the life expectancy of breast cancer patients. Our studies demonstrate that TNBC cells surviving the chronic treatment of chemotherapeutic drugs show significantly higher expression of the dual serine/threonine and tyrosine protein kinase (DSTYK) than non-treated parental cells. In our in vitro cellular models, DSTYK knockout via the CRISPR/Cas9-mediated technique results in apoptotic cell death of chemoresistant cells upon drug treatment. Moreover, DSTYK knockout promotes chemotherapeutic drug-induced tumor cell death in an orthotopic mouse model. These findings suggest that DSTYK exerts an important and previously unknown role in promoting chemoresistance. Our studies provide fundamental insight into the role of DSTYK in chemoresistance in TNBC cells and lay the foundation for the development of new strategies targeting DSTYK for improving TNBC therapy.

Keywords: CRISPR/Cas9; DSTYK; TNBC; breast cancer; chemoresistance.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Statistical analysis of DSTYK-related survival probability in breast cancer patients. (A) Survival curves of breast cancer patients with differential DSTYK expression published in The Human Protein Atlas (HPA). Patients include all four cancer stages and those stages are not available (n/a). The survival probabilities were generated and are publicly available at website (www.proteinatlas.org, accessed on 14 December 2021). (B) Survival curves of TNBC patients and (C) TNBC patients only treated with chemotherapy from the Kaplan-Meier Plotter dataset statistical analysis (https://kmplot.com/analysis/index.php?p=service&cancer=breast, accessed on 14 December 2021).
Figure 2
Figure 2
Schematic diagram of the procedure for generating chemoresistant TNBC cells. (A) SUM102PT and MDA-MB-468 cells were exposed to DOX/DXL. The concentration of drugs was gradually increased (DOX/DXL:1 µM/10 nM, 2 µM/20 nM, 4 µM/40 nM, 6 µM/60 nM, 8 µM/80 nM, 10 µM/100 nM) until stable proliferation at 10 µM/100 nM was established. (B) MTT assays to compare chemoresistance between SUM102PT and SUM102PTR cells, and (C) between MDA-MB-468 and MDA-MB-468R cells. The data presented represent the mean values ± SD (n = 3). (D) TUNEL apoptosis assays comparing the chemoresistance between SUM102PT and SUM102PTR, and between MDA-MB-468 and MDA-MB-468R cells. (E) Representative images from flow cytometry analyses to compare the levels of cell death by staining with cell surface apoptotic marker annexin V and the necrotic marker 7-ADD between parental cells and resistant SUM102PT cells treated with or without DOX/DXL (2 µM/20 nM) or MDA-MB-468 cells treated with or without DOX/DXL (4 µM/40 nM). (F) The quantification of the results from (E). The data represent mean values ± SD; ** p < 0.01; *** p < 0.001.
Figure 3
Figure 3
DSTYK expression correlates with chemoresistance. (A) The mRNA level of DSTYK between parental and chemoresistant cells by reverse-transcription PCR (RT-PCR) and (B) qPCR. (C,D) WB and quantification to detect the expressions of DSTYK in SUM102PT and SUM102PTR cells, and MDA-MB-468 and MDA-MB-468R cells, respectively. Hsp90 was used as the loading control. Experiments were repeated three times; ** p < 0.01; one-way ANOVA.
Figure 4
Figure 4
DSTYK is essential to the chemoresistance in TNBC cells. (A) MTT assays to compare the chemoresistance treated with different concentrations of DOX/DXL. The data presented represents the mean values ± SD (n = 3). The levels of cellular DSTYK protein are indicated by the included WB. Hsp90 was used as the loading control. (B) WBs to detect the apoptosis marker cleaved-caspase 3 before and after DOX/DXL treatment (0.5/5 μM/nM for SUM102PTR cells; 1/10 μM/nM for MDA-MB-468R cells). GAPDH was used as the loading control. (C–F) Quantification analyses of (B). (G) Clonogenic assays to confirm that chemoresistant cells regain chemosensitivity after DSTYKKO. DOX/DXL 4 μM/40 nm for SUM102PTR cells and 6 μM/60 nm for MDA-MB-468R cells. (H,I) MTT assays to compare the chemoresistance among parental control cells, parental DSTYKKO cells, and parental DSTYKKO-RE cells. Experiments were repeated three times; ns, not significant; * p < 0.05; **: p < 0.01; *** p < 0.001, one-way ANOVA.
Figure 5
Figure 5
DSTYKKO sensitizes chemoresistant cells to chemotherapy drug treatment. (A) Representative images from flow cytometry analyses to compare the levels of cell surface apoptotic marker annexin V and the necrotic marker 7-ADD between control and DSTYKKO groups in SUM102PTR cells or MDA-MB-468R cells with or without DOX/DXL. DOX/DXL 2 μM/20 nm for SUM102PTR cells and 4 μM/40 nm for MDA-MB-468R cells. (B) The data in (A) are quantified. For each cell line, the left panel is the result for non-drug treated and the right panel is the result for drug-treated. The data represents mean values ± SD; ns, not significant; ** p < 0.01; *** p < 0.001. (C,D) DSTYKKO resistant cell growth for 5 days and total numbers of cells were counted on days 1, 3, and 5 (n = 3 independent experiments) in SUM102PTR cells and MDA-MB-468R cells.
Figure 6
Figure 6
DSTYK promotes chemoresistance in vivo. (A) Representative images of one set of tumors derived from control and DSTYKKO SUM102PTR cells. (B) The graph represents the tumor growth volume observed for the different groups (as labeled) derived from SUM102PTR cells. The data represents mean values ± SD. (C) Quantification of tumors after excision and being weighed at the end point; n = 7; n.s., not significant; * p < 0.05; *** p < 0.001, one-way ANOVA. (D) Representative images of one set of tumors derived from control and DSTYKKO MDA-MB-468R cells. (E) The graph represents the tumor growth volume observed for the different groups derived from MDA-MB-468R cells (as labeled) until reaching the endpoint. The data represents mean values ± SD. (F) Quantification of tumors after excision and weighing at the end point; n = 7; n.s., not significant; * p < 0.05; *** p < 0.001, one-way ANOVA.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Ji X., Lu Y., Tian H., Meng X., Wei M., Cho W.C. Chemoresistance mechanisms of breast cancer and their countermeasures. Biomed. Pharmacother. 2019;114:108800. doi: 10.1016/j.biopha.2019.108800. - DOI - PubMed
    1. Bordoloi D., Roy N.K., Monisha J., Padmavathi G., Kunnumakkara A.B. Multi-Targeted Agents in Cancer Cell Chemosensitization: What We Learnt from Curcumin Thus Far. Recent Pat. Anticancer Drug Discov. 2016;11:67–97. doi: 10.2174/1574892810666151020101706. - DOI - PubMed
    1. da Silva J.L., Cardoso Nunes N.C., Izetti P., de Mesquita G.G., de Melo A.C. Triple negative breast cancer: A thorough review of biomarkers. Crit. Rev. Oncol. Hematol. 2020;145:102855. doi: 10.1016/j.critrevonc.2019.102855. - DOI - PubMed
    1. Bianchini G., Balko J.M., Mayer I.A., Sanders M.E., Gianni L. Triple-negative breast cancer: Challenges and opportunities of a heterogeneous disease. Nat. Rev. Clin. Oncol. 2016;13:674–690. doi: 10.1038/nrclinonc.2016.66. - DOI - PMC - PubMed
    1. Omarini C., Guaitoli G., Pipitone S., Moscetti L., Cortesi L., Cascinu S., Piacentini F. Neoadjuvant treatments in triple-negative breast cancer patients: Where we are now and where we are going. Cancer Manag. Res. 2018;10:91–103. doi: 10.2147/CMAR.S146658. - DOI - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources