Limits...
Molecular signatures of sanguinarine in human pancreatic cancer cells: A large scale label-free comparative proteomics approach.

Singh CK, Kaur S, George J, Nihal M, Pellitteri Hahn MC, Scarlett CO, Ahmad N - Oncotarget (2015)

Bottom Line: Further validation by qRT-PCR and immunoblot analyses demonstrated that the dual specificity phosphatase-4 (DUSP4) was significantly upregulated by sanguinarine in BxPC-3 and MIA PaCa-2 cells.Sanguinarine treatment also caused down-regulation of HIF1α and PCNA, and increased cleavage of PARP and Caspase-7.Taken together, sanguinarine appears to have pleotropic effects, as it modulates multiple key signaling pathways, supporting the potential usefulness of sanguinarine against pancreatic cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Wisconsin, Madison, WI, USA.

ABSTRACT
Pancreatic cancer remains one of the most lethal of all human malignancies with its incidence nearly equaling its mortality rate. Therefore, it's crucial to identify newer mechanism-based agents and targets to effectively manage pancreatic cancer. Plant-derived agents/drugs have historically been useful in cancer therapeutics. Sanguinarine is a plant alkaloid with anti-proliferative effects against cancers, including pancreatic cancer. This study was designed to determine the mechanism of sanguinarine's effects in pancreatic cancer with a hope to obtain useful information to improve the therapeutic options for the management of this neoplasm. We employed a quantitative proteomics approach to define the mechanism of sanguinarine's effects in human pancreatic cancer cells. Proteins from control and sanguinarine-treated pancreatic cancer cells were digested with trypsin, run by nano-LC/MS/MS, and identified with the help of Swiss-Prot database. Results from replicate injections were processed with the SIEVE software to identify proteins with differential expression. We identified 37 differentially expressed proteins (from a total of 3107), which are known to be involved in variety of cellular processes. Four of these proteins (IL33, CUL5, GPS1 and DUSP4) appear to occupy regulatory nodes in key pathways. Further validation by qRT-PCR and immunoblot analyses demonstrated that the dual specificity phosphatase-4 (DUSP4) was significantly upregulated by sanguinarine in BxPC-3 and MIA PaCa-2 cells. Sanguinarine treatment also caused down-regulation of HIF1α and PCNA, and increased cleavage of PARP and Caspase-7. Taken together, sanguinarine appears to have pleotropic effects, as it modulates multiple key signaling pathways, supporting the potential usefulness of sanguinarine against pancreatic cancer.

No MeSH data available.


Related in: MedlinePlus

IPA analysis of proteins changing in abundance with sanguinarine treatment(A) Association of canonical signaling pathways with modulated proteins are shown. The proteins which demonstrated significant change (95% confidence interval with statistical significance) were subjected to IPA analysis. The top 26 canonical pathways were identified as significantly altered upon sanguinarine treatment. The line bar represents the threshold of significance ( p = 0.05). (B) IPA was further used to categorize the proteins on the basis of disease and/or functional relation to the altered proteins.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4496359&req=5

Figure 3: IPA analysis of proteins changing in abundance with sanguinarine treatment(A) Association of canonical signaling pathways with modulated proteins are shown. The proteins which demonstrated significant change (95% confidence interval with statistical significance) were subjected to IPA analysis. The top 26 canonical pathways were identified as significantly altered upon sanguinarine treatment. The line bar represents the threshold of significance ( p = 0.05). (B) IPA was further used to categorize the proteins on the basis of disease and/or functional relation to the altered proteins.

Mentions: The relationships, putative networks and canonical pathways analysis of differentially expressed proteins were performed by Ingenuity Pathway Analysis (IPA) (Ingenuity Inc.). The selected 37 proteins (Table 1) were uploaded to the IPA module with their corresponding Swiss-Prot IDs and respective fold changes to map proteins into biological networks and to retrieve functions and key pathways. We were able to identify association of 26 canonical pathways with these selected proteins. Among these, the protein ubiquitination pathway which generally involves in post translational modification, appears the top hit. The protein ubiquitination system functions in a variety of cellular processes, including apoptosis, cell cycle and division, DNA transcription and repair, response to stress and extracellular modulators. Further, as shown in Figure 3A, we identified an involvement of several important signaling pathways in the biological response of sanguinarine.


Molecular signatures of sanguinarine in human pancreatic cancer cells: A large scale label-free comparative proteomics approach.

Singh CK, Kaur S, George J, Nihal M, Pellitteri Hahn MC, Scarlett CO, Ahmad N - Oncotarget (2015)

IPA analysis of proteins changing in abundance with sanguinarine treatment(A) Association of canonical signaling pathways with modulated proteins are shown. The proteins which demonstrated significant change (95% confidence interval with statistical significance) were subjected to IPA analysis. The top 26 canonical pathways were identified as significantly altered upon sanguinarine treatment. The line bar represents the threshold of significance ( p = 0.05). (B) IPA was further used to categorize the proteins on the basis of disease and/or functional relation to the altered proteins.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496359&req=5

Figure 3: IPA analysis of proteins changing in abundance with sanguinarine treatment(A) Association of canonical signaling pathways with modulated proteins are shown. The proteins which demonstrated significant change (95% confidence interval with statistical significance) were subjected to IPA analysis. The top 26 canonical pathways were identified as significantly altered upon sanguinarine treatment. The line bar represents the threshold of significance ( p = 0.05). (B) IPA was further used to categorize the proteins on the basis of disease and/or functional relation to the altered proteins.
Mentions: The relationships, putative networks and canonical pathways analysis of differentially expressed proteins were performed by Ingenuity Pathway Analysis (IPA) (Ingenuity Inc.). The selected 37 proteins (Table 1) were uploaded to the IPA module with their corresponding Swiss-Prot IDs and respective fold changes to map proteins into biological networks and to retrieve functions and key pathways. We were able to identify association of 26 canonical pathways with these selected proteins. Among these, the protein ubiquitination pathway which generally involves in post translational modification, appears the top hit. The protein ubiquitination system functions in a variety of cellular processes, including apoptosis, cell cycle and division, DNA transcription and repair, response to stress and extracellular modulators. Further, as shown in Figure 3A, we identified an involvement of several important signaling pathways in the biological response of sanguinarine.

Bottom Line: Further validation by qRT-PCR and immunoblot analyses demonstrated that the dual specificity phosphatase-4 (DUSP4) was significantly upregulated by sanguinarine in BxPC-3 and MIA PaCa-2 cells.Sanguinarine treatment also caused down-regulation of HIF1α and PCNA, and increased cleavage of PARP and Caspase-7.Taken together, sanguinarine appears to have pleotropic effects, as it modulates multiple key signaling pathways, supporting the potential usefulness of sanguinarine against pancreatic cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Wisconsin, Madison, WI, USA.

ABSTRACT
Pancreatic cancer remains one of the most lethal of all human malignancies with its incidence nearly equaling its mortality rate. Therefore, it's crucial to identify newer mechanism-based agents and targets to effectively manage pancreatic cancer. Plant-derived agents/drugs have historically been useful in cancer therapeutics. Sanguinarine is a plant alkaloid with anti-proliferative effects against cancers, including pancreatic cancer. This study was designed to determine the mechanism of sanguinarine's effects in pancreatic cancer with a hope to obtain useful information to improve the therapeutic options for the management of this neoplasm. We employed a quantitative proteomics approach to define the mechanism of sanguinarine's effects in human pancreatic cancer cells. Proteins from control and sanguinarine-treated pancreatic cancer cells were digested with trypsin, run by nano-LC/MS/MS, and identified with the help of Swiss-Prot database. Results from replicate injections were processed with the SIEVE software to identify proteins with differential expression. We identified 37 differentially expressed proteins (from a total of 3107), which are known to be involved in variety of cellular processes. Four of these proteins (IL33, CUL5, GPS1 and DUSP4) appear to occupy regulatory nodes in key pathways. Further validation by qRT-PCR and immunoblot analyses demonstrated that the dual specificity phosphatase-4 (DUSP4) was significantly upregulated by sanguinarine in BxPC-3 and MIA PaCa-2 cells. Sanguinarine treatment also caused down-regulation of HIF1α and PCNA, and increased cleavage of PARP and Caspase-7. Taken together, sanguinarine appears to have pleotropic effects, as it modulates multiple key signaling pathways, supporting the potential usefulness of sanguinarine against pancreatic cancer.

No MeSH data available.


Related in: MedlinePlus