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Quercetin induces cell cycle arrest and apoptosis in CD133(+) cancer stem cells of human colorectal HT29 cancer cell line and enhances anticancer effects of doxorubicin.

Atashpour S, Fouladdel S, Movahhed TK, Barzegar E, Ghahremani MH, Ostad SN, Azizi E - Iran J Basic Med Sci (2015)

Bottom Line: Quercetin has anticancer effects with the advantage of exhibiting low side effects.Therefore, we evaluated the anticancer effects of quercetin and doxorubicin (Dox) in HT29 cancer cells and its isolated CD133(+) CSCs.The CSCs were a minor population with a significantly high level of drug resistance within the HT29 cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Molecular Research Lab, Department of Pharmacology and Toxicology, Tehran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Objectives: The colorectal cancer stem cells (CSCs) with the CD133(+) phenotype are a rare fraction of cancer cells with the ability of self-renewal, unlimited proliferation and resistance to treatment. Quercetin has anticancer effects with the advantage of exhibiting low side effects. Therefore, we evaluated the anticancer effects of quercetin and doxorubicin (Dox) in HT29 cancer cells and its isolated CD133(+) CSCs.

Materials and methods: The CSCs from HT29 cells were isolated using CD133 antibody conjugated to magnetic beads by MACS. Anticancer effects of quercetin and Dox alone and in combination on HT29 cells and CSCs were evaluated using MTT cytotoxicity assay and flow cytometry analysis of cell cycle distribution and apoptosis induction.

Results: The CD133(+) CSCs comprised about 10% of HT29 cells. Quercetin and Dox alone and in combination inhibited cell proliferation and induced apoptosis in HT29 cells and to a lesser extent in CSCs. Quercetin enhanced cytotoxicity and apoptosis induction of Dox at low concentration in both cell populations. Quercetin and Dox and their combination induced G2/M arrest in the HT29 cells and to a lesser extent in CSCs.

Conclusion: The CSCs were a minor population with a significantly high level of drug resistance within the HT29 cancer cells. Quercetin alone exhibited significant cytotoxic effects on HT29 cells and also increased cytoxicity of Dox in combination therapy. Altogether, our data showed that adding quercetin to Dox chemotherapy is an effective strategy for treatment of both CSCs and bulk tumor cells.

No MeSH data available.


Related in: MedlinePlus

Cell cycle alteration in HT29 cell line and its isolated CD133+ cancer stem cells. The HT29 cancer cells (A) and its isolated CD133+ cancer stem cells (B) were treated with Dox and Quer alone and in combination for 72 hr to determine cell cycle distribution pattern using DAPI staining by flow cytometry analysis. Data are presented as the mean±SE of three independent experiments. # denotes P<0.01 and *, P<0.001 for significant difference between treatments in comparison to control RPMI. Dox: Doxorubicin; Quer: Quercetin
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Figure 5: Cell cycle alteration in HT29 cell line and its isolated CD133+ cancer stem cells. The HT29 cancer cells (A) and its isolated CD133+ cancer stem cells (B) were treated with Dox and Quer alone and in combination for 72 hr to determine cell cycle distribution pattern using DAPI staining by flow cytometry analysis. Data are presented as the mean±SE of three independent experiments. # denotes P<0.01 and *, P<0.001 for significant difference between treatments in comparison to control RPMI. Dox: Doxorubicin; Quer: Quercetin

Mentions: In order to understand whether the growth inhibitory effect of Quer and Dox was due to cell cycle arrest, we evaluated the effects of treatments on cell cycle distribution using DAPI staining and flow cytometry analysis. In HT29 cell line, 55.42% of control (RPMI) treated cells were in G0/G1, 23.55% in S and 21% in G2/M phases of cell cycle (Figure 5A). Flow cytometry analysis revealed that Dox treatment at IC50 induced accumulation of HT29 cells in G2/M phase (60.7%) compared to the control cells (21%). Quer treatment at IC50 also induced G2/M arrest (60%) in HT29 cells almost the same as Dox treatment. Importantly, similar level of G2/M arrest (64%) was observed in HT29 cells treated with combination of Dox and Quer at much lower concentration than their IC50 (Figure 5A). Importantly, the pattern of cell cycle distribution of the isolated CD133+ CSCs showed significantly higher percentage of cells at G0/G1 (79%) in comparison to parental HT29 cells (55%) in control (RPMI) condition (Figure 5A and B). This further indicated that CSCs were more in quiescent phase and therefore, less responsive to Dox treatment at IC50, which resulted in significantly less accumulation of CSCs in G2/M phase (33.79%) in comparison to parental HT29 cells (60.7%). Similar results for G2/M arrest (35.96%) were observed following Quer treatment at IC50 or combination of Dox and Quer (40.48%) at much lower concentration than their IC50 (Figure 5B).


Quercetin induces cell cycle arrest and apoptosis in CD133(+) cancer stem cells of human colorectal HT29 cancer cell line and enhances anticancer effects of doxorubicin.

Atashpour S, Fouladdel S, Movahhed TK, Barzegar E, Ghahremani MH, Ostad SN, Azizi E - Iran J Basic Med Sci (2015)

Cell cycle alteration in HT29 cell line and its isolated CD133+ cancer stem cells. The HT29 cancer cells (A) and its isolated CD133+ cancer stem cells (B) were treated with Dox and Quer alone and in combination for 72 hr to determine cell cycle distribution pattern using DAPI staining by flow cytometry analysis. Data are presented as the mean±SE of three independent experiments. # denotes P<0.01 and *, P<0.001 for significant difference between treatments in comparison to control RPMI. Dox: Doxorubicin; Quer: Quercetin
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556754&req=5

Figure 5: Cell cycle alteration in HT29 cell line and its isolated CD133+ cancer stem cells. The HT29 cancer cells (A) and its isolated CD133+ cancer stem cells (B) were treated with Dox and Quer alone and in combination for 72 hr to determine cell cycle distribution pattern using DAPI staining by flow cytometry analysis. Data are presented as the mean±SE of three independent experiments. # denotes P<0.01 and *, P<0.001 for significant difference between treatments in comparison to control RPMI. Dox: Doxorubicin; Quer: Quercetin
Mentions: In order to understand whether the growth inhibitory effect of Quer and Dox was due to cell cycle arrest, we evaluated the effects of treatments on cell cycle distribution using DAPI staining and flow cytometry analysis. In HT29 cell line, 55.42% of control (RPMI) treated cells were in G0/G1, 23.55% in S and 21% in G2/M phases of cell cycle (Figure 5A). Flow cytometry analysis revealed that Dox treatment at IC50 induced accumulation of HT29 cells in G2/M phase (60.7%) compared to the control cells (21%). Quer treatment at IC50 also induced G2/M arrest (60%) in HT29 cells almost the same as Dox treatment. Importantly, similar level of G2/M arrest (64%) was observed in HT29 cells treated with combination of Dox and Quer at much lower concentration than their IC50 (Figure 5A). Importantly, the pattern of cell cycle distribution of the isolated CD133+ CSCs showed significantly higher percentage of cells at G0/G1 (79%) in comparison to parental HT29 cells (55%) in control (RPMI) condition (Figure 5A and B). This further indicated that CSCs were more in quiescent phase and therefore, less responsive to Dox treatment at IC50, which resulted in significantly less accumulation of CSCs in G2/M phase (33.79%) in comparison to parental HT29 cells (60.7%). Similar results for G2/M arrest (35.96%) were observed following Quer treatment at IC50 or combination of Dox and Quer (40.48%) at much lower concentration than their IC50 (Figure 5B).

Bottom Line: Quercetin has anticancer effects with the advantage of exhibiting low side effects.Therefore, we evaluated the anticancer effects of quercetin and doxorubicin (Dox) in HT29 cancer cells and its isolated CD133(+) CSCs.The CSCs were a minor population with a significantly high level of drug resistance within the HT29 cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Molecular Research Lab, Department of Pharmacology and Toxicology, Tehran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Objectives: The colorectal cancer stem cells (CSCs) with the CD133(+) phenotype are a rare fraction of cancer cells with the ability of self-renewal, unlimited proliferation and resistance to treatment. Quercetin has anticancer effects with the advantage of exhibiting low side effects. Therefore, we evaluated the anticancer effects of quercetin and doxorubicin (Dox) in HT29 cancer cells and its isolated CD133(+) CSCs.

Materials and methods: The CSCs from HT29 cells were isolated using CD133 antibody conjugated to magnetic beads by MACS. Anticancer effects of quercetin and Dox alone and in combination on HT29 cells and CSCs were evaluated using MTT cytotoxicity assay and flow cytometry analysis of cell cycle distribution and apoptosis induction.

Results: The CD133(+) CSCs comprised about 10% of HT29 cells. Quercetin and Dox alone and in combination inhibited cell proliferation and induced apoptosis in HT29 cells and to a lesser extent in CSCs. Quercetin enhanced cytotoxicity and apoptosis induction of Dox at low concentration in both cell populations. Quercetin and Dox and their combination induced G2/M arrest in the HT29 cells and to a lesser extent in CSCs.

Conclusion: The CSCs were a minor population with a significantly high level of drug resistance within the HT29 cancer cells. Quercetin alone exhibited significant cytotoxic effects on HT29 cells and also increased cytoxicity of Dox in combination therapy. Altogether, our data showed that adding quercetin to Dox chemotherapy is an effective strategy for treatment of both CSCs and bulk tumor cells.

No MeSH data available.


Related in: MedlinePlus