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Combined genomic and phenotype screening reveals secretory factor SPINK1 as an invasion and survival factor associated with patient prognosis in breast cancer.

Soon WW, Miller LD, Black MA, Dalmasso C, Chan XB, Pang B, Ong CW, Salto-Tellez M, Desai KV, Liu ET - EMBO Mol Med (2011)

Bottom Line: In ER- cases, the prognostic effect did not reach statistical significance.Intriguingly, these anti-apoptotic effects of SPINK1 were abrogated by mutations of its protease inhibition domain.Because SPINK1 effects are abrogated by neutralizing antibodies, we suggest that SPINK1 is a viable potential therapeutic target in breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Cancer Biology and Pharmacology, Genome Institute of Singapore.

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Related in: MedlinePlus

Proliferation rescue assay of SPINK1 in MDA-MB-231Incorporation of BrDU in MDA-MB-231 cells upon SPINK1-kd, and rescue with bioactive SPINK1-CM. Significance of knockdown was measured against scrambled control, while significance of SPINK1CM was measured against VecCM as a control (*p < 0.005).
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fig06: Proliferation rescue assay of SPINK1 in MDA-MB-231Incorporation of BrDU in MDA-MB-231 cells upon SPINK1-kd, and rescue with bioactive SPINK1-CM. Significance of knockdown was measured against scrambled control, while significance of SPINK1CM was measured against VecCM as a control (*p < 0.005).

Mentions: To determine the specificity of our SPINK1 siRNA constructs by a phenotypic rescue, we generated two sources of recombinant SPINK1 by over-expressing SPINK1 in MCF-7 and SF9 insect cells (Fig S5, panels A–C of Supporting information). Conditioned media (CM) was collected from both sources (referred to as SPINK1 CM and SF9spink1 CM), as well as from cells transfected with empty vector as controls (vector-CM and SF9vecCM, respectively). Treatment of C2siRNA cells with SPINK1-CM resulted in 5-bromodeoxyuridine (BrDU) incorporation showing that these cells survived and could subsequently divide (Fig 6). The control Vec-CM was unable to rescue this cell death. This confirms the specificity of the siRNA used as well as the functionality of the recombinant SPINK1. Interestingly, neither MCF-7 cells over-expressing SPINK1 (MCF-7 SPINK1OE) nor various breast cancer cell lines treated with SPINK1-CM showed increased proliferation (Fig S6 of Supporting information). This further supports the notion that SPINK1 is probably an important survival factor rather than a potent growth inducer in breast cancer.


Combined genomic and phenotype screening reveals secretory factor SPINK1 as an invasion and survival factor associated with patient prognosis in breast cancer.

Soon WW, Miller LD, Black MA, Dalmasso C, Chan XB, Pang B, Ong CW, Salto-Tellez M, Desai KV, Liu ET - EMBO Mol Med (2011)

Proliferation rescue assay of SPINK1 in MDA-MB-231Incorporation of BrDU in MDA-MB-231 cells upon SPINK1-kd, and rescue with bioactive SPINK1-CM. Significance of knockdown was measured against scrambled control, while significance of SPINK1CM was measured against VecCM as a control (*p < 0.005).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3377086&req=5

fig06: Proliferation rescue assay of SPINK1 in MDA-MB-231Incorporation of BrDU in MDA-MB-231 cells upon SPINK1-kd, and rescue with bioactive SPINK1-CM. Significance of knockdown was measured against scrambled control, while significance of SPINK1CM was measured against VecCM as a control (*p < 0.005).
Mentions: To determine the specificity of our SPINK1 siRNA constructs by a phenotypic rescue, we generated two sources of recombinant SPINK1 by over-expressing SPINK1 in MCF-7 and SF9 insect cells (Fig S5, panels A–C of Supporting information). Conditioned media (CM) was collected from both sources (referred to as SPINK1 CM and SF9spink1 CM), as well as from cells transfected with empty vector as controls (vector-CM and SF9vecCM, respectively). Treatment of C2siRNA cells with SPINK1-CM resulted in 5-bromodeoxyuridine (BrDU) incorporation showing that these cells survived and could subsequently divide (Fig 6). The control Vec-CM was unable to rescue this cell death. This confirms the specificity of the siRNA used as well as the functionality of the recombinant SPINK1. Interestingly, neither MCF-7 cells over-expressing SPINK1 (MCF-7 SPINK1OE) nor various breast cancer cell lines treated with SPINK1-CM showed increased proliferation (Fig S6 of Supporting information). This further supports the notion that SPINK1 is probably an important survival factor rather than a potent growth inducer in breast cancer.

Bottom Line: In ER- cases, the prognostic effect did not reach statistical significance.Intriguingly, these anti-apoptotic effects of SPINK1 were abrogated by mutations of its protease inhibition domain.Because SPINK1 effects are abrogated by neutralizing antibodies, we suggest that SPINK1 is a viable potential therapeutic target in breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Cancer Biology and Pharmacology, Genome Institute of Singapore.

Show MeSH
Related in: MedlinePlus