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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

Apoptosis induction 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 evaluate induction of apoptosis using Annexin V-FITC/PI double-staining by flow cytometry analysis. The percentage of early and late apoptotic cells were presented for each treatment group. Data are presented as the mean±SE of three independent experiments. *denotes P<0.001 for significant difference between treatments in comparison to control RPMI. Dox: Doxorubicin; Quer: Quercetin
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Figure 6: Apoptosis induction 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 evaluate induction of apoptosis using Annexin V-FITC/PI double-staining by flow cytometry analysis. The percentage of early and late apoptotic cells were presented for each treatment group. Data are presented as the mean±SE of three independent experiments. *denotes P<0.001 for significant difference between treatments in comparison to control RPMI. Dox: Doxorubicin; Quer: Quercetin

Mentions: In order to study the induction of apoptosis, HT29 cells and its isolated CD133+ CSCs were treated with IC50 of Dox and Quer alone and in combination. Then cells were double-stained with AnnexinV-FITC and PI and analyzed by flow cytometry. Flow cytometry analysis data were processed with FloMax software to determine the percentage of necrotic (Q1: AnnexinV-FITC-/PI+), late apoptotic (Q2: AnnexinV-FITC+/PI+), viable (Q3: AnnexinV-FITC-/PI-), and early apoptotic (Q4: AnnexinV-FITC+/PI-) cells. The sums of percentages of early (Q4) and late (Q2) apoptotic cells in HT29 cells treated with IC50 of Dox (44.98%) and Quer (49.7%) alone and in combination (56.77%) were significantly greater than that of control cells (0.26%) (Figure 6A).


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)

Apoptosis induction 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 evaluate induction of apoptosis using Annexin V-FITC/PI double-staining by flow cytometry analysis. The percentage of early and late apoptotic cells were presented for each treatment group. Data are presented as the mean±SE of three independent experiments. *denotes 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 6: Apoptosis induction 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 evaluate induction of apoptosis using Annexin V-FITC/PI double-staining by flow cytometry analysis. The percentage of early and late apoptotic cells were presented for each treatment group. Data are presented as the mean±SE of three independent experiments. *denotes P<0.001 for significant difference between treatments in comparison to control RPMI. Dox: Doxorubicin; Quer: Quercetin
Mentions: In order to study the induction of apoptosis, HT29 cells and its isolated CD133+ CSCs were treated with IC50 of Dox and Quer alone and in combination. Then cells were double-stained with AnnexinV-FITC and PI and analyzed by flow cytometry. Flow cytometry analysis data were processed with FloMax software to determine the percentage of necrotic (Q1: AnnexinV-FITC-/PI+), late apoptotic (Q2: AnnexinV-FITC+/PI+), viable (Q3: AnnexinV-FITC-/PI-), and early apoptotic (Q4: AnnexinV-FITC+/PI-) cells. The sums of percentages of early (Q4) and late (Q2) apoptotic cells in HT29 cells treated with IC50 of Dox (44.98%) and Quer (49.7%) alone and in combination (56.77%) were significantly greater than that of control cells (0.26%) (Figure 6A).

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