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Effect of Thymoquinone on P53 Gene Expression and Consequence Apoptosis in Breast Cancer Cell Line.

Dastjerdi MN, Mehdiabady EM, Iranpour FG, Bahramian H - Int J Prev Med (2016)

Bottom Line: The IC50 value for the TQ in MCF-7 cells was 25 μM that determined using MTT assay.The flow cytometry and real-time PCR results showed that TQ could induce apoptosis in MCF-7 cells, and the P53 gene expression was dramatically up-regulated by ascending time, respectively.Hence, there was significant difference in 48 and 72 h.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomical Sciences and Molecular Biology, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran.

ABSTRACT

Background: Nigella sativa has been a nutritional flavoring factor and natural treatment for many ailments for so many years in medical science. Earlier studies have been reported that thymoquinone (TQ), an active compound of its seed, contains anticancer properties. Previous studies have shown that TQ induces apoptosis in breast cancer cells but it is unclear the role of P53 in the apoptotic pathway. Hereby, this study reports the potency of TQ on expression of tumor suppressor gene P53 and apoptosis induction in breast cancer cell line Michigan Cancer Foundation-7 (MCF-7).

Methods: MCF-7 cell line was cultured and treated with TQ, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out for evaluating the half-maximal inhibitory concentration (IC50) values after 24 h of treatment. The percentage of apoptotic cells was measured by flow cytometry. Real-time polymerase chain reaction (PCR) was performed to estimate the messenger RNA expression of P53 in MCF-7 cell line at different times.

Results: The IC50 value for the TQ in MCF-7 cells was 25 μM that determined using MTT assay. The flow cytometry and real-time PCR results showed that TQ could induce apoptosis in MCF-7 cells, and the P53 gene expression was dramatically up-regulated by ascending time, respectively. Hence, there was significant difference in 48 and 72 h.

Conclusions: Our results demonstrated that TQ could induce apoptosis in MCF-7 cells through up-regulation of P53 expression in breast cancer cell line (MCF-7) by time-dependent manner.

No MeSH data available.


Related in: MedlinePlus

Half-maximal inhibitory concentration assay for half-maximal inhibitory concentration analysis of thymoquinone in Michigan Cancer Foundation-7 cancer cell lines after 24 h of treatment. Cells were incubated with or without the thymoquinone using 25 μM dose. The relative amount of viable cells was estimated by measuring the absorbance of the cell suspension after incubation with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A graph of viability versus drug concentration was used to calculate half-maximal inhibitory concentration values for Michigan Cancer Foundation-7 cell line
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Figure 1: Half-maximal inhibitory concentration assay for half-maximal inhibitory concentration analysis of thymoquinone in Michigan Cancer Foundation-7 cancer cell lines after 24 h of treatment. Cells were incubated with or without the thymoquinone using 25 μM dose. The relative amount of viable cells was estimated by measuring the absorbance of the cell suspension after incubation with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A graph of viability versus drug concentration was used to calculate half-maximal inhibitory concentration values for Michigan Cancer Foundation-7 cell line

Mentions: After the treatment of MCF-7 cells with MTT solution in this study, the dark blue formazan crystals were seen in cells, which indicated their metabolic activity. The reduction in the number of cells was dependent on the cell type as shown by the IC50 index. The IC50 value for the TQ was established. The results showed that the essential TQ concentration to achieve the IC50 in MCF-7 cells at 24 h was 25 μmol/L [Figure 1].


Effect of Thymoquinone on P53 Gene Expression and Consequence Apoptosis in Breast Cancer Cell Line.

Dastjerdi MN, Mehdiabady EM, Iranpour FG, Bahramian H - Int J Prev Med (2016)

Half-maximal inhibitory concentration assay for half-maximal inhibitory concentration analysis of thymoquinone in Michigan Cancer Foundation-7 cancer cell lines after 24 h of treatment. Cells were incubated with or without the thymoquinone using 25 μM dose. The relative amount of viable cells was estimated by measuring the absorbance of the cell suspension after incubation with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A graph of viability versus drug concentration was used to calculate half-maximal inhibitory concentration values for Michigan Cancer Foundation-7 cell line
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4837800&req=5

Figure 1: Half-maximal inhibitory concentration assay for half-maximal inhibitory concentration analysis of thymoquinone in Michigan Cancer Foundation-7 cancer cell lines after 24 h of treatment. Cells were incubated with or without the thymoquinone using 25 μM dose. The relative amount of viable cells was estimated by measuring the absorbance of the cell suspension after incubation with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A graph of viability versus drug concentration was used to calculate half-maximal inhibitory concentration values for Michigan Cancer Foundation-7 cell line
Mentions: After the treatment of MCF-7 cells with MTT solution in this study, the dark blue formazan crystals were seen in cells, which indicated their metabolic activity. The reduction in the number of cells was dependent on the cell type as shown by the IC50 index. The IC50 value for the TQ was established. The results showed that the essential TQ concentration to achieve the IC50 in MCF-7 cells at 24 h was 25 μmol/L [Figure 1].

Bottom Line: The IC50 value for the TQ in MCF-7 cells was 25 μM that determined using MTT assay.The flow cytometry and real-time PCR results showed that TQ could induce apoptosis in MCF-7 cells, and the P53 gene expression was dramatically up-regulated by ascending time, respectively.Hence, there was significant difference in 48 and 72 h.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomical Sciences and Molecular Biology, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran.

ABSTRACT

Background: Nigella sativa has been a nutritional flavoring factor and natural treatment for many ailments for so many years in medical science. Earlier studies have been reported that thymoquinone (TQ), an active compound of its seed, contains anticancer properties. Previous studies have shown that TQ induces apoptosis in breast cancer cells but it is unclear the role of P53 in the apoptotic pathway. Hereby, this study reports the potency of TQ on expression of tumor suppressor gene P53 and apoptosis induction in breast cancer cell line Michigan Cancer Foundation-7 (MCF-7).

Methods: MCF-7 cell line was cultured and treated with TQ, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out for evaluating the half-maximal inhibitory concentration (IC50) values after 24 h of treatment. The percentage of apoptotic cells was measured by flow cytometry. Real-time polymerase chain reaction (PCR) was performed to estimate the messenger RNA expression of P53 in MCF-7 cell line at different times.

Results: The IC50 value for the TQ in MCF-7 cells was 25 μM that determined using MTT assay. The flow cytometry and real-time PCR results showed that TQ could induce apoptosis in MCF-7 cells, and the P53 gene expression was dramatically up-regulated by ascending time, respectively. Hence, there was significant difference in 48 and 72 h.

Conclusions: Our results demonstrated that TQ could induce apoptosis in MCF-7 cells through up-regulation of P53 expression in breast cancer cell line (MCF-7) by time-dependent manner.

No MeSH data available.


Related in: MedlinePlus