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Midostaurin preferentially attenuates proliferation of triple-negative breast cancer cell lines through inhibition of Aurora kinase family.

Kawai M, Nakashima A, Kamada S, Kikkawa U - J. Biomed. Sci. (2015)

Bottom Line: It is, thus, required to develop an effective therapeutic reagent to treat TNBC.Following studies indicated that midostaurin attenuates the phosphorylation reaction mediated by Aurora kinase in the cells and directly inhibits this protein kinase in vitro, and that this reagent induces apoptosis accompanying accumulation of 4N and 8N DNA cells in TNBC cells.The precise study of midostaurin on cell growth will contribute to the development of the drug for the treatment of TNBC.

View Article: PubMed Central - PubMed

Affiliation: Biosignal Research Center, Kobe University, Kobe, 657-8501, Japan. 090s381s@stu.kobe-u.ac.jp.

ABSTRACT

Background: Breast cancer is classified into three subtypes by the expression of biomarker receptors such as hormone receptors and human epidermal growth factor receptor 2. Triple-negative breast cancer (TNBC) expresses none of these receptors and has an aggressive phenotype with a poor prognosis, which is insensitive to the drugs that target the hormone receptors and human epidermal growth factor receptor 2. It is, thus, required to develop an effective therapeutic reagent to treat TNBC.

Results: The study using a panel of 19 breast cancer cell lines revealed that midostaurin, a multi-target protein kinase inhibitor, suppresses preferentially the growth of TNBC cells comparing with non-TNBC cells. Clustering analysis of the drug activity data for the panel of cancer cell lines predicted that midostaurin shares the target with Aurora kinase inhibitors. Following studies indicated that midostaurin attenuates the phosphorylation reaction mediated by Aurora kinase in the cells and directly inhibits this protein kinase in vitro, and that this reagent induces apoptosis accompanying accumulation of 4N and 8N DNA cells in TNBC cells.

Conclusion: Midostaurin suppresses the proliferation of TNBC cells among the breast cancer cell lines presumably through the inhibition of the Aurora kinase family. The precise study of midostaurin on cell growth will contribute to the development of the drug for the treatment of TNBC.

No MeSH data available.


Related in: MedlinePlus

Western blot analysis of breast cancer cell lines. Cell lysates from the 19 breast cancer cell lines were subjected to Western blot analysis. Aurora kinases A and B, p-Aurora kinase A and B, and Histone H3 and p-Histone H3 Ser10 were detected using respective antibodies. The experiments were carried out after calibration by using GAPDH as an internal control. Breast cancer subtypes are indicated as follows: gray, TNBC; light gray, HER2; white, ER+
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Fig4: Western blot analysis of breast cancer cell lines. Cell lysates from the 19 breast cancer cell lines were subjected to Western blot analysis. Aurora kinases A and B, p-Aurora kinase A and B, and Histone H3 and p-Histone H3 Ser10 were detected using respective antibodies. The experiments were carried out after calibration by using GAPDH as an internal control. Breast cancer subtypes are indicated as follows: gray, TNBC; light gray, HER2; white, ER+

Mentions: To determine the role of Aurora kinases in TNBC, the expression level of Aurora kinases A and B was examined in the cancer cell lines (Fig. 4). Expression of Aurora kinases A and B was detected among various cell lines, and was generally high in the TNBC cell lines relative to the non-TNBC cell lines. The phosphorylation of Aurora kinases A and B at Thr288 and Thr232, respectively, as well as that of Histone H3 at Ser10 was evaluated an indicator of the Aurora kinase activity [32]. Notably, the phosphorylation of Histone H3 at Ser10, recognized by Aurora kinase B [32], was high in most of the TNBC cell lines in agreement with the expression level of Aurora kinases A and B. The phosphorylation of Aurora kinases A and B was significantly high in two TNBC cell lines, HCC1806 and MDA-MB-468, but was weak in other cell lines. Thus, the effect of midostaurin was examined on these phosphorylation reactions in the TNBC cell line of MDA-MB-468 (Fig. 5a). The midostaurin treatment significantly reduced the Aurora kinase-mediated phosphorylation reactions in the cell line. Although the effect was weaker than that of VX-680, midostaurin at 1 μM attenuated the phosphorylation of Aurora kinases A and B as well as Histone H3. In vitro kinase assay showed that midostaurin at 1 μM inhibited the kinase activity of GST-tagged Aurora kinases A and B by 16 and 34 %, respectively (Fig. 5b). Furthermore, the negative correlation was observed between the phosphorylation level of Histone H3 Ser10 and cell viability after the midostaurin treatment (Fig. 6a): Pearson’s correlation coefficient was −0.51 (p = 0.021 by Pearson correlation test). Most of the TNBC cell lines showed a high level of phosphorylation of Histone H3 Ser10 with low cell viability. The phosphorylation level of Histone H3 was higher in TNBC cell lines than that in non-TNBC cell lines (Fig. 6b). These results indicate that midostaurin directly inhibits Aurora kinases A and B, and subsequently reduces cell viability predominantly in the TNBC cells.Fig. 4


Midostaurin preferentially attenuates proliferation of triple-negative breast cancer cell lines through inhibition of Aurora kinase family.

Kawai M, Nakashima A, Kamada S, Kikkawa U - J. Biomed. Sci. (2015)

Western blot analysis of breast cancer cell lines. Cell lysates from the 19 breast cancer cell lines were subjected to Western blot analysis. Aurora kinases A and B, p-Aurora kinase A and B, and Histone H3 and p-Histone H3 Ser10 were detected using respective antibodies. The experiments were carried out after calibration by using GAPDH as an internal control. Breast cancer subtypes are indicated as follows: gray, TNBC; light gray, HER2; white, ER+
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4491224&req=5

Fig4: Western blot analysis of breast cancer cell lines. Cell lysates from the 19 breast cancer cell lines were subjected to Western blot analysis. Aurora kinases A and B, p-Aurora kinase A and B, and Histone H3 and p-Histone H3 Ser10 were detected using respective antibodies. The experiments were carried out after calibration by using GAPDH as an internal control. Breast cancer subtypes are indicated as follows: gray, TNBC; light gray, HER2; white, ER+
Mentions: To determine the role of Aurora kinases in TNBC, the expression level of Aurora kinases A and B was examined in the cancer cell lines (Fig. 4). Expression of Aurora kinases A and B was detected among various cell lines, and was generally high in the TNBC cell lines relative to the non-TNBC cell lines. The phosphorylation of Aurora kinases A and B at Thr288 and Thr232, respectively, as well as that of Histone H3 at Ser10 was evaluated an indicator of the Aurora kinase activity [32]. Notably, the phosphorylation of Histone H3 at Ser10, recognized by Aurora kinase B [32], was high in most of the TNBC cell lines in agreement with the expression level of Aurora kinases A and B. The phosphorylation of Aurora kinases A and B was significantly high in two TNBC cell lines, HCC1806 and MDA-MB-468, but was weak in other cell lines. Thus, the effect of midostaurin was examined on these phosphorylation reactions in the TNBC cell line of MDA-MB-468 (Fig. 5a). The midostaurin treatment significantly reduced the Aurora kinase-mediated phosphorylation reactions in the cell line. Although the effect was weaker than that of VX-680, midostaurin at 1 μM attenuated the phosphorylation of Aurora kinases A and B as well as Histone H3. In vitro kinase assay showed that midostaurin at 1 μM inhibited the kinase activity of GST-tagged Aurora kinases A and B by 16 and 34 %, respectively (Fig. 5b). Furthermore, the negative correlation was observed between the phosphorylation level of Histone H3 Ser10 and cell viability after the midostaurin treatment (Fig. 6a): Pearson’s correlation coefficient was −0.51 (p = 0.021 by Pearson correlation test). Most of the TNBC cell lines showed a high level of phosphorylation of Histone H3 Ser10 with low cell viability. The phosphorylation level of Histone H3 was higher in TNBC cell lines than that in non-TNBC cell lines (Fig. 6b). These results indicate that midostaurin directly inhibits Aurora kinases A and B, and subsequently reduces cell viability predominantly in the TNBC cells.Fig. 4

Bottom Line: It is, thus, required to develop an effective therapeutic reagent to treat TNBC.Following studies indicated that midostaurin attenuates the phosphorylation reaction mediated by Aurora kinase in the cells and directly inhibits this protein kinase in vitro, and that this reagent induces apoptosis accompanying accumulation of 4N and 8N DNA cells in TNBC cells.The precise study of midostaurin on cell growth will contribute to the development of the drug for the treatment of TNBC.

View Article: PubMed Central - PubMed

Affiliation: Biosignal Research Center, Kobe University, Kobe, 657-8501, Japan. 090s381s@stu.kobe-u.ac.jp.

ABSTRACT

Background: Breast cancer is classified into three subtypes by the expression of biomarker receptors such as hormone receptors and human epidermal growth factor receptor 2. Triple-negative breast cancer (TNBC) expresses none of these receptors and has an aggressive phenotype with a poor prognosis, which is insensitive to the drugs that target the hormone receptors and human epidermal growth factor receptor 2. It is, thus, required to develop an effective therapeutic reagent to treat TNBC.

Results: The study using a panel of 19 breast cancer cell lines revealed that midostaurin, a multi-target protein kinase inhibitor, suppresses preferentially the growth of TNBC cells comparing with non-TNBC cells. Clustering analysis of the drug activity data for the panel of cancer cell lines predicted that midostaurin shares the target with Aurora kinase inhibitors. Following studies indicated that midostaurin attenuates the phosphorylation reaction mediated by Aurora kinase in the cells and directly inhibits this protein kinase in vitro, and that this reagent induces apoptosis accompanying accumulation of 4N and 8N DNA cells in TNBC cells.

Conclusion: Midostaurin suppresses the proliferation of TNBC cells among the breast cancer cell lines presumably through the inhibition of the Aurora kinase family. The precise study of midostaurin on cell growth will contribute to the development of the drug for the treatment of TNBC.

No MeSH data available.


Related in: MedlinePlus