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Inhibition of SHP2 in basal-like and triple-negative breast cells induces basal-to-luminal transition, hormone dependency, and sensitivity to anti-hormone treatment.

Zhao H, Agazie YM - BMC Cancer (2015)

Bottom Line: The Src homology phosphotyrosyl phosphatase 2 (SHP2) is a positive effector of cell growth and survival signaling as well transformation induced by multiple tyrosine kinase oncogenes.The occurrence of BLT was confirmed by the loss of the basal marker alpha smooth muscle actin and the acquisition of the luminal marker cytokeratin 18 (CK18) expression.Our data show that inhibition of SHP2 induces BLT, ERα expression, dependency on estrogen for growth, and sensitivity to anti-hormone therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and The Marry Babb Randolph Cancer Center School of Medicine, West Virginia University, Morgantown, WV, 26506, USA. ttangela168@gmail.com.

ABSTRACT

Background: The Src homology phosphotyrosyl phosphatase 2 (SHP2) is a positive effector of cell growth and survival signaling as well transformation induced by multiple tyrosine kinase oncogenes. Since the basal-like and triple-negative breast cancer (BTBC) is characterized by dysregulation of multiple tyrosine kinase oncogenes, we wanted to determine the importance of SHP2 in BTBC cell lines.

Methods: Short hairpin RNA-based and dominant-negative expression-based SHP2 inhibition techniques were used to interrogate the functional importance of SHP2 in BTBC cell biology. In addition, cell viability and proliferation assays were used to determine hormone dependency for growth and sensitivity to anti-estrogen treatment.

Results: We show that inhibition of SHP2 in BTBC cells induces luminal-like epithelial morphology while suppressing the mesenchymal and invasive property. We have termed this process as basal-to-luminal transition (BLT). The occurrence of BLT was confirmed by the loss of the basal marker alpha smooth muscle actin and the acquisition of the luminal marker cytokeratin 18 (CK18) expression. Furthermore, the occurrence of BLT led to estrogen receptor alpha (ERα) expression, hormone dependency, and sensitivity to tamoxifen treatment.

Conclusions: Our data show that inhibition of SHP2 induces BLT, ERα expression, dependency on estrogen for growth, and sensitivity to anti-hormone therapy. Therefore, inhibition of SHP2 may provide a therapeutic benefit in basal-like and triple-negative breast cancer.

No MeSH data available.


Related in: MedlinePlus

Effect of SHP2 silencing on cell migration. Silencing SHP2 expression retards cell migration in the MDA-MB231 (A) and the MDA-MB468 (B) cells. Data shown was from the control and the shRNA-2 cells derived from the respective cell lines. C) Effect of SHP2 silencing on the ability of the MDA-MB231 cells to degrade FITC-labeled collagen in 3D matrigel invasion assay. D) Fluorescent intensity measurement of collagen degradation by the MDA-MB231 cells. E) Effect of SHP2 silencing on the ability of the MDA-MB468 cells to degrade FITC-labeled collagen in 3D matrigel invasion assay. F) Fluorescent intensity measurement of collagen degradation by the MDA-MB468 cells. Data shown was mean ± SEM of three independent experiments.
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Fig2: Effect of SHP2 silencing on cell migration. Silencing SHP2 expression retards cell migration in the MDA-MB231 (A) and the MDA-MB468 (B) cells. Data shown was from the control and the shRNA-2 cells derived from the respective cell lines. C) Effect of SHP2 silencing on the ability of the MDA-MB231 cells to degrade FITC-labeled collagen in 3D matrigel invasion assay. D) Fluorescent intensity measurement of collagen degradation by the MDA-MB231 cells. E) Effect of SHP2 silencing on the ability of the MDA-MB468 cells to degrade FITC-labeled collagen in 3D matrigel invasion assay. F) Fluorescent intensity measurement of collagen degradation by the MDA-MB468 cells. Data shown was mean ± SEM of three independent experiments.

Mentions: Most BTBC cell lines, including the cells used in this study are highly mesenchymal and exhibit enhanced migratory and invasive properties [29]. The morphological changes induced by SHP2 inhibition were indicative of the importance of SHP2 in these properties. To test this possibility, we employed the simple wound-healing assay in which cells will be induced to migrate and fill the space created by scratching. In agreement with our previous report on effect of SHP2 in cell polarity [18], silencing SHP2 expression retarded cell migration. While the control cells were able to heal the wound in 24 hours, the SHP2-silenced cells were unable to do so even in 48 hours (Figure 2A and B). Similar results were obtained when SHP2 function was inhibited by expression of C459S-SHP2 (Additional file 2: Figure S2A and B). Hence, SHP2 is essential for the migratory behavior of BTBC cells.Figure 2


Inhibition of SHP2 in basal-like and triple-negative breast cells induces basal-to-luminal transition, hormone dependency, and sensitivity to anti-hormone treatment.

Zhao H, Agazie YM - BMC Cancer (2015)

Effect of SHP2 silencing on cell migration. Silencing SHP2 expression retards cell migration in the MDA-MB231 (A) and the MDA-MB468 (B) cells. Data shown was from the control and the shRNA-2 cells derived from the respective cell lines. C) Effect of SHP2 silencing on the ability of the MDA-MB231 cells to degrade FITC-labeled collagen in 3D matrigel invasion assay. D) Fluorescent intensity measurement of collagen degradation by the MDA-MB231 cells. E) Effect of SHP2 silencing on the ability of the MDA-MB468 cells to degrade FITC-labeled collagen in 3D matrigel invasion assay. F) Fluorescent intensity measurement of collagen degradation by the MDA-MB468 cells. Data shown was mean ± SEM of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359540&req=5

Fig2: Effect of SHP2 silencing on cell migration. Silencing SHP2 expression retards cell migration in the MDA-MB231 (A) and the MDA-MB468 (B) cells. Data shown was from the control and the shRNA-2 cells derived from the respective cell lines. C) Effect of SHP2 silencing on the ability of the MDA-MB231 cells to degrade FITC-labeled collagen in 3D matrigel invasion assay. D) Fluorescent intensity measurement of collagen degradation by the MDA-MB231 cells. E) Effect of SHP2 silencing on the ability of the MDA-MB468 cells to degrade FITC-labeled collagen in 3D matrigel invasion assay. F) Fluorescent intensity measurement of collagen degradation by the MDA-MB468 cells. Data shown was mean ± SEM of three independent experiments.
Mentions: Most BTBC cell lines, including the cells used in this study are highly mesenchymal and exhibit enhanced migratory and invasive properties [29]. The morphological changes induced by SHP2 inhibition were indicative of the importance of SHP2 in these properties. To test this possibility, we employed the simple wound-healing assay in which cells will be induced to migrate and fill the space created by scratching. In agreement with our previous report on effect of SHP2 in cell polarity [18], silencing SHP2 expression retarded cell migration. While the control cells were able to heal the wound in 24 hours, the SHP2-silenced cells were unable to do so even in 48 hours (Figure 2A and B). Similar results were obtained when SHP2 function was inhibited by expression of C459S-SHP2 (Additional file 2: Figure S2A and B). Hence, SHP2 is essential for the migratory behavior of BTBC cells.Figure 2

Bottom Line: The Src homology phosphotyrosyl phosphatase 2 (SHP2) is a positive effector of cell growth and survival signaling as well transformation induced by multiple tyrosine kinase oncogenes.The occurrence of BLT was confirmed by the loss of the basal marker alpha smooth muscle actin and the acquisition of the luminal marker cytokeratin 18 (CK18) expression.Our data show that inhibition of SHP2 induces BLT, ERα expression, dependency on estrogen for growth, and sensitivity to anti-hormone therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and The Marry Babb Randolph Cancer Center School of Medicine, West Virginia University, Morgantown, WV, 26506, USA. ttangela168@gmail.com.

ABSTRACT

Background: The Src homology phosphotyrosyl phosphatase 2 (SHP2) is a positive effector of cell growth and survival signaling as well transformation induced by multiple tyrosine kinase oncogenes. Since the basal-like and triple-negative breast cancer (BTBC) is characterized by dysregulation of multiple tyrosine kinase oncogenes, we wanted to determine the importance of SHP2 in BTBC cell lines.

Methods: Short hairpin RNA-based and dominant-negative expression-based SHP2 inhibition techniques were used to interrogate the functional importance of SHP2 in BTBC cell biology. In addition, cell viability and proliferation assays were used to determine hormone dependency for growth and sensitivity to anti-estrogen treatment.

Results: We show that inhibition of SHP2 in BTBC cells induces luminal-like epithelial morphology while suppressing the mesenchymal and invasive property. We have termed this process as basal-to-luminal transition (BLT). The occurrence of BLT was confirmed by the loss of the basal marker alpha smooth muscle actin and the acquisition of the luminal marker cytokeratin 18 (CK18) expression. Furthermore, the occurrence of BLT led to estrogen receptor alpha (ERα) expression, hormone dependency, and sensitivity to tamoxifen treatment.

Conclusions: Our data show that inhibition of SHP2 induces BLT, ERα expression, dependency on estrogen for growth, and sensitivity to anti-hormone therapy. Therefore, inhibition of SHP2 may provide a therapeutic benefit in basal-like and triple-negative breast cancer.

No MeSH data available.


Related in: MedlinePlus