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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

(a) Relative levels of apoptotic cells in Michigan Cancer Foundation-7 treated with thymoquinone for different times. Cells incubated with the vehicle dimethyl sulfoxide were used as a control. The percentage of apoptotic cells was measured using the Annexin V-FITC flow cytometry 1-histogram and propidium iodide flow cytometry 2-histogram. Dunnett's test was used for considered comparisons (*P < 0.05) (in any figure, right upper quarter show late apoptotic cells, left upper quarter show necrosis cells, right lower quarter show early apoptotic cells and left lower quarter show normal cells). (b) Cells that are Annexin V-positive and propidium iodide negative are in early apoptosis as phosphatidyl serine translocation has occurred; although, the plasma membrane remains intact. Cells that are positive for both Annexin V and propidium iodide either are in the late stages of apoptosis or are already dead as phosphatidyl serine translocation has occurred and the loss of plasma membrane integrity is visible. *P < 0.05 compared to controls. Dunnett's test was used for considered comparisons (*P < 0.05)
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Figure 2: (a) Relative levels of apoptotic cells in Michigan Cancer Foundation-7 treated with thymoquinone for different times. Cells incubated with the vehicle dimethyl sulfoxide were used as a control. The percentage of apoptotic cells was measured using the Annexin V-FITC flow cytometry 1-histogram and propidium iodide flow cytometry 2-histogram. Dunnett's test was used for considered comparisons (*P < 0.05) (in any figure, right upper quarter show late apoptotic cells, left upper quarter show necrosis cells, right lower quarter show early apoptotic cells and left lower quarter show normal cells). (b) Cells that are Annexin V-positive and propidium iodide negative are in early apoptosis as phosphatidyl serine translocation has occurred; although, the plasma membrane remains intact. Cells that are positive for both Annexin V and propidium iodide either are in the late stages of apoptosis or are already dead as phosphatidyl serine translocation has occurred and the loss of plasma membrane integrity is visible. *P < 0.05 compared to controls. Dunnett's test was used for considered comparisons (*P < 0.05)

Mentions: To establish the anti-apoptosis potential of the TQ, we first investigated the effects of TQ on the proliferation in MCF-7 cells. The flow cytometry results showed that TQ at different time points (24, 48 and 72 h) could induce apoptosis in MCF-7 cells, and it was increased with ascending time. TQ arrested MCF-7 cells proliferation (40% of inhibition) in 72 h and (approximately 30% of inhibition) in 48 h [Figure 2a and b, P < 0.05]. DMSO was used in the control sample (vehicle drugs). Using Dunnett test, average of apoptotic cells at different times were compared with the control group. Groups 48 h and 72 h had significant difference to the control group (*P < 0.05).


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)

(a) Relative levels of apoptotic cells in Michigan Cancer Foundation-7 treated with thymoquinone for different times. Cells incubated with the vehicle dimethyl sulfoxide were used as a control. The percentage of apoptotic cells was measured using the Annexin V-FITC flow cytometry 1-histogram and propidium iodide flow cytometry 2-histogram. Dunnett's test was used for considered comparisons (*P < 0.05) (in any figure, right upper quarter show late apoptotic cells, left upper quarter show necrosis cells, right lower quarter show early apoptotic cells and left lower quarter show normal cells). (b) Cells that are Annexin V-positive and propidium iodide negative are in early apoptosis as phosphatidyl serine translocation has occurred; although, the plasma membrane remains intact. Cells that are positive for both Annexin V and propidium iodide either are in the late stages of apoptosis or are already dead as phosphatidyl serine translocation has occurred and the loss of plasma membrane integrity is visible. *P < 0.05 compared to controls. Dunnett's test was used for considered comparisons (*P < 0.05)
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: (a) Relative levels of apoptotic cells in Michigan Cancer Foundation-7 treated with thymoquinone for different times. Cells incubated with the vehicle dimethyl sulfoxide were used as a control. The percentage of apoptotic cells was measured using the Annexin V-FITC flow cytometry 1-histogram and propidium iodide flow cytometry 2-histogram. Dunnett's test was used for considered comparisons (*P < 0.05) (in any figure, right upper quarter show late apoptotic cells, left upper quarter show necrosis cells, right lower quarter show early apoptotic cells and left lower quarter show normal cells). (b) Cells that are Annexin V-positive and propidium iodide negative are in early apoptosis as phosphatidyl serine translocation has occurred; although, the plasma membrane remains intact. Cells that are positive for both Annexin V and propidium iodide either are in the late stages of apoptosis or are already dead as phosphatidyl serine translocation has occurred and the loss of plasma membrane integrity is visible. *P < 0.05 compared to controls. Dunnett's test was used for considered comparisons (*P < 0.05)
Mentions: To establish the anti-apoptosis potential of the TQ, we first investigated the effects of TQ on the proliferation in MCF-7 cells. The flow cytometry results showed that TQ at different time points (24, 48 and 72 h) could induce apoptosis in MCF-7 cells, and it was increased with ascending time. TQ arrested MCF-7 cells proliferation (40% of inhibition) in 72 h and (approximately 30% of inhibition) in 48 h [Figure 2a and b, P < 0.05]. DMSO was used in the control sample (vehicle drugs). Using Dunnett test, average of apoptotic cells at different times were compared with the control group. Groups 48 h and 72 h had significant difference to the control group (*P < 0.05).

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