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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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Cell responsiveness of colorectal adenocarcinoma cell lines after treatment with Ciglitazone.2D-PAGE gels were performed in (A) HT29 and (B) SW480 cells after treatment with 5 µM Ciglitazone. (1) Fluorescence scans of untreated cells, (2) fluorescence scans of cells treated with Ciglitazone, (3) autoradiography scans of untreated cells, (4) autoradiography scans of cells treated with Ciglitazone. Cell cycle distribution of (C) HT29 and (D) SW480 cells treated with increasing concentrations of Ciglitazone.
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pone-0114158-g003: Cell responsiveness of colorectal adenocarcinoma cell lines after treatment with Ciglitazone.2D-PAGE gels were performed in (A) HT29 and (B) SW480 cells after treatment with 5 µM Ciglitazone. (1) Fluorescence scans of untreated cells, (2) fluorescence scans of cells treated with Ciglitazone, (3) autoradiography scans of untreated cells, (4) autoradiography scans of cells treated with Ciglitazone. Cell cycle distribution of (C) HT29 and (D) SW480 cells treated with increasing concentrations of Ciglitazone.

Mentions: Protein amounts determined by fluorescence detection were hardly affected upon treatment with CIG at 5 µM (Fig. 3A and 3B upper panel). Not a single protein was found up- or downregulated more than three-fold by CIG treatment in a significant manner. Since most proteins detected by fluorescence are relatively abundant, measurement of total protein amounts was not sensitive enough to assess the cells' responsiveness to CIG. In order to compare cell responsiveness to drug treatment in a more sensitive fashion we performed autoradiography scans [26], [27]. Cells treated with CIG at 5 µM responded with an upregulation of protein synthesis as compared to cells incubated with DMSO. In HT29 cells the synthesis was increased by 91±40% versus 74±26% in SW480 cells, indicating a slightly higher responsiveness of HT29 cells.


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)

Cell responsiveness of colorectal adenocarcinoma cell lines after treatment with Ciglitazone.2D-PAGE gels were performed in (A) HT29 and (B) SW480 cells after treatment with 5 µM Ciglitazone. (1) Fluorescence scans of untreated cells, (2) fluorescence scans of cells treated with Ciglitazone, (3) autoradiography scans of untreated cells, (4) autoradiography scans of cells treated with Ciglitazone. Cell cycle distribution of (C) HT29 and (D) SW480 cells treated with increasing concentrations of Ciglitazone.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0114158-g003: Cell responsiveness of colorectal adenocarcinoma cell lines after treatment with Ciglitazone.2D-PAGE gels were performed in (A) HT29 and (B) SW480 cells after treatment with 5 µM Ciglitazone. (1) Fluorescence scans of untreated cells, (2) fluorescence scans of cells treated with Ciglitazone, (3) autoradiography scans of untreated cells, (4) autoradiography scans of cells treated with Ciglitazone. Cell cycle distribution of (C) HT29 and (D) SW480 cells treated with increasing concentrations of Ciglitazone.
Mentions: Protein amounts determined by fluorescence detection were hardly affected upon treatment with CIG at 5 µM (Fig. 3A and 3B upper panel). Not a single protein was found up- or downregulated more than three-fold by CIG treatment in a significant manner. Since most proteins detected by fluorescence are relatively abundant, measurement of total protein amounts was not sensitive enough to assess the cells' responsiveness to CIG. In order to compare cell responsiveness to drug treatment in a more sensitive fashion we performed autoradiography scans [26], [27]. Cells treated with CIG at 5 µM responded with an upregulation of protein synthesis as compared to cells incubated with DMSO. In HT29 cells the synthesis was increased by 91±40% versus 74±26% in SW480 cells, indicating a slightly higher responsiveness of HT29 cells.

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