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Autonomous inhibition of apoptosis correlates with responsiveness of colon carcinoma cell lines to ciglitazone.

Baron DM, Kaindl U, Haudek-Prinz VJ, Bayer E, Röhrl C, Gerner C, Marian B - PLoS ONE (2014)

Bottom Line: Resistance to therapy is common and often results in patients succumbing to the disease.To verify the data we performed shotgun analysis using the same treatment procedure as in 2D-experiments.Biological functions of the identified proteins were mainly associated with apoptosis regulation, chaperoning, intrinsic inflammation, and DNA repair.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine I, Clinic for Internal Medicine I, Institute of Cancer Research, Medical University of Vienna, Vienna, Austria; Department of Anesthesia, General Intensive Care, and Pain Management Medical, Medical University of Vienna, Vienna, Austria.

ABSTRACT
Colorectal cancer is a leading cause of mortality worldwide. Resistance to therapy is common and often results in patients succumbing to the disease. The mechanisms of resistance are poorly understood. Cells basically have two possibilities to survive a treatment with potentially apoptosis-inducing substances. They can make use of their existing proteins to counteract the induced reactions or quickly upregulate protective factors to evade the apoptotic signal. To identify protein patterns involved in resistance to apoptosis, we studied two colorectal adenocarcinoma cell lines with different growth responses to low-molar concentrations of the thiazolidinedione Ciglitazone: HT29 cells underwent apoptosis, whereas SW480 cells increased cell number. Fluorescence detection and autoradiography scans of 2D-PAGE gels were performed in both cell lines to assess protein synthesis and turnover, respectively. To verify the data we performed shotgun analysis using the same treatment procedure as in 2D-experiments. Biological functions of the identified proteins were mainly associated with apoptosis regulation, chaperoning, intrinsic inflammation, and DNA repair. The present study suggests that different growth response of two colorectal carcinoma cell lines after treatment with Ciglitazone results from cell-specific protein synthesis and differences in protein regulation.

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Protein synthesis in untreated colorectal adenocarcinoma cell lines.Representative 2D-PAGE gels of untreated (A) HT29 and (B) SW480 cells. Proteins synthesized to a greater extent in HT29 cells are indicated by hexagons, those in SW480 cells by circles. Accession numbers of proteins listed in Table 1 are annotated according to the UniProtKB/Swiss-Prot database. (C) Total proteins identified by shotgun analysis in HT29 and SW480 cells.
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pone-0114158-g002: Protein synthesis in untreated colorectal adenocarcinoma cell lines.Representative 2D-PAGE gels of untreated (A) HT29 and (B) SW480 cells. Proteins synthesized to a greater extent in HT29 cells are indicated by hexagons, those in SW480 cells by circles. Accession numbers of proteins listed in Table 1 are annotated according to the UniProtKB/Swiss-Prot database. (C) Total proteins identified by shotgun analysis in HT29 and SW480 cells.

Mentions: Therefore, our first goal was to compare protein synthesis patterns between untreated cell lines in order to recognize proteins, which are constantly expressed by the cells and might explain survival-promoting effects. In order to elucidate whether baseline alterations in protein synthesis might be associated with the different responsiveness to treatment with CIG, 2D-PAGE with fluorescence detection was performed in untreated HT29 and SW480 cell lines. Overall, 5 cytoplasmic proteins were synthesized at least three-fold higher in HT29 cells than in SW480 cells (p<0.05, Table 1 and Fig. 2A). In comparison, 12 cytoplasmic proteins were synthetized at least three-fold higher in SW480 cells than in HT29 cells (p<0.05, Table 1 and Fig. 2B), whereas 570 proteins were identified in both cell lines with less than three-fold difference. Gel-to-gel variations, however, pose an important limitation for proteomic research. When tissue culture, cell preparation, and 2D-gel-runs are replicated independently slight variations cannot be avoided [36].


Autonomous inhibition of apoptosis correlates with responsiveness of colon carcinoma cell lines to ciglitazone.

Baron DM, Kaindl U, Haudek-Prinz VJ, Bayer E, Röhrl C, Gerner C, Marian B - PLoS ONE (2014)

Protein synthesis in untreated colorectal adenocarcinoma cell lines.Representative 2D-PAGE gels of untreated (A) HT29 and (B) SW480 cells. Proteins synthesized to a greater extent in HT29 cells are indicated by hexagons, those in SW480 cells by circles. Accession numbers of proteins listed in Table 1 are annotated according to the UniProtKB/Swiss-Prot database. (C) Total proteins identified by shotgun analysis in HT29 and SW480 cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263530&req=5

pone-0114158-g002: Protein synthesis in untreated colorectal adenocarcinoma cell lines.Representative 2D-PAGE gels of untreated (A) HT29 and (B) SW480 cells. Proteins synthesized to a greater extent in HT29 cells are indicated by hexagons, those in SW480 cells by circles. Accession numbers of proteins listed in Table 1 are annotated according to the UniProtKB/Swiss-Prot database. (C) Total proteins identified by shotgun analysis in HT29 and SW480 cells.
Mentions: Therefore, our first goal was to compare protein synthesis patterns between untreated cell lines in order to recognize proteins, which are constantly expressed by the cells and might explain survival-promoting effects. In order to elucidate whether baseline alterations in protein synthesis might be associated with the different responsiveness to treatment with CIG, 2D-PAGE with fluorescence detection was performed in untreated HT29 and SW480 cell lines. Overall, 5 cytoplasmic proteins were synthesized at least three-fold higher in HT29 cells than in SW480 cells (p<0.05, Table 1 and Fig. 2A). In comparison, 12 cytoplasmic proteins were synthetized at least three-fold higher in SW480 cells than in HT29 cells (p<0.05, Table 1 and Fig. 2B), whereas 570 proteins were identified in both cell lines with less than three-fold difference. Gel-to-gel variations, however, pose an important limitation for proteomic research. When tissue culture, cell preparation, and 2D-gel-runs are replicated independently slight variations cannot be avoided [36].

Bottom Line: Resistance to therapy is common and often results in patients succumbing to the disease.To verify the data we performed shotgun analysis using the same treatment procedure as in 2D-experiments.Biological functions of the identified proteins were mainly associated with apoptosis regulation, chaperoning, intrinsic inflammation, and DNA repair.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine I, Clinic for Internal Medicine I, Institute of Cancer Research, Medical University of Vienna, Vienna, Austria; Department of Anesthesia, General Intensive Care, and Pain Management Medical, Medical University of Vienna, Vienna, Austria.

ABSTRACT
Colorectal cancer is a leading cause of mortality worldwide. Resistance to therapy is common and often results in patients succumbing to the disease. The mechanisms of resistance are poorly understood. Cells basically have two possibilities to survive a treatment with potentially apoptosis-inducing substances. They can make use of their existing proteins to counteract the induced reactions or quickly upregulate protective factors to evade the apoptotic signal. To identify protein patterns involved in resistance to apoptosis, we studied two colorectal adenocarcinoma cell lines with different growth responses to low-molar concentrations of the thiazolidinedione Ciglitazone: HT29 cells underwent apoptosis, whereas SW480 cells increased cell number. Fluorescence detection and autoradiography scans of 2D-PAGE gels were performed in both cell lines to assess protein synthesis and turnover, respectively. To verify the data we performed shotgun analysis using the same treatment procedure as in 2D-experiments. Biological functions of the identified proteins were mainly associated with apoptosis regulation, chaperoning, intrinsic inflammation, and DNA repair. The present study suggests that different growth response of two colorectal carcinoma cell lines after treatment with Ciglitazone results from cell-specific protein synthesis and differences in protein regulation.

Show MeSH
Related in: MedlinePlus