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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

Silencing SHP2 expression induces acini-like structure formation. A) Light pictures of parental, control and SHP2-silenced MDA-MB231 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. B) Light pictures of parental, control and SHP2-silenced MDA-MB468 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. Pictures of acini-like structures formed by the MCF-10A cells cultured under identical conditions is shown in both A and B for comparison. C) Effect of SHP2 silencing on expression of basal and luminal markers and estrogen receptor alpha (ERα). Data presented is representative of at least three independent experiments.
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Fig4: Silencing SHP2 expression induces acini-like structure formation. A) Light pictures of parental, control and SHP2-silenced MDA-MB231 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. B) Light pictures of parental, control and SHP2-silenced MDA-MB468 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. Pictures of acini-like structures formed by the MCF-10A cells cultured under identical conditions is shown in both A and B for comparison. C) Effect of SHP2 silencing on expression of basal and luminal markers and estrogen receptor alpha (ERα). Data presented is representative of at least three independent experiments.

Mentions: As shown in Figure 1 and Additional file 1: Figure S1, silencing SHP2 expression induced a luminal-like epithelial morphology in BTBC cells. Based on these findings, we reasoned that inhibition of SHP2 may induce conversion of BTBC cells to a luminal lineage, which we termed basal-to-luminal transition (BLT). This possibility was tested by the laminin-rich basement membrane (LRBM) culture where non-transformed luminal breast epithelial cells like the MCF10A form acini-like structures while breast cancer cells produce a continuously-growing amorphous cellular mass [19,23]. As expected, the control cells formed a disorganized cellular mass that continued to grow and invade, whereas the SHP2-silenced cells grew very slowly, forming acini-like spheroid structures that are comparable to those formed by the control MCF-10A cells (Figure 4A and B). These results suggest that inhibition of SHP2 reverses the transformation and invasive phenotype and induces BLT in BTBC cells.Figure 4


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)

Silencing SHP2 expression induces acini-like structure formation. A) Light pictures of parental, control and SHP2-silenced MDA-MB231 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. B) Light pictures of parental, control and SHP2-silenced MDA-MB468 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. Pictures of acini-like structures formed by the MCF-10A cells cultured under identical conditions is shown in both A and B for comparison. C) Effect of SHP2 silencing on expression of basal and luminal markers and estrogen receptor alpha (ERα). Data presented is representative of at least three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: Silencing SHP2 expression induces acini-like structure formation. A) Light pictures of parental, control and SHP2-silenced MDA-MB231 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. B) Light pictures of parental, control and SHP2-silenced MDA-MB468 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. Pictures of acini-like structures formed by the MCF-10A cells cultured under identical conditions is shown in both A and B for comparison. C) Effect of SHP2 silencing on expression of basal and luminal markers and estrogen receptor alpha (ERα). Data presented is representative of at least three independent experiments.
Mentions: As shown in Figure 1 and Additional file 1: Figure S1, silencing SHP2 expression induced a luminal-like epithelial morphology in BTBC cells. Based on these findings, we reasoned that inhibition of SHP2 may induce conversion of BTBC cells to a luminal lineage, which we termed basal-to-luminal transition (BLT). This possibility was tested by the laminin-rich basement membrane (LRBM) culture where non-transformed luminal breast epithelial cells like the MCF10A form acini-like structures while breast cancer cells produce a continuously-growing amorphous cellular mass [19,23]. As expected, the control cells formed a disorganized cellular mass that continued to grow and invade, whereas the SHP2-silenced cells grew very slowly, forming acini-like spheroid structures that are comparable to those formed by the control MCF-10A cells (Figure 4A and B). These results suggest that inhibition of SHP2 reverses the transformation and invasive phenotype and induces BLT in BTBC cells.Figure 4

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