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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

Results of real-time quantitative polymerase chain reaction before and after thymoquinone treatment at different times on the P53 messenger RNA expression in Michigan Cancer Foundation-7 cell. Relative expression levels were obtained using the comparative Ct (ΔΔCt) method (*P < 0.05)
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Figure 3: Results of real-time quantitative polymerase chain reaction before and after thymoquinone treatment at different times on the P53 messenger RNA expression in Michigan Cancer Foundation-7 cell. Relative expression levels were obtained using the comparative Ct (ΔΔCt) method (*P < 0.05)

Mentions: It was suggested that apoptotic induction in cancer cells by TQ requires the activation of P53 gene expression. To examine this hypothesis, we used MCF-7 cell line, as cancerous cell line. We examined the inhibitory effects of 25 μmol/L TQ (based on IC50 index) at different times on the mRNA expression of P53 in MCF-7 cell line. The P53 gene expression was dramatically up-regulated by ascending time, in particular, at 72 h treatment its expression was increased significantly [Figure 3, P < 0.05]. Using Dunnett test, average of P53 gene expression at different times was 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)

Results of real-time quantitative polymerase chain reaction before and after thymoquinone treatment at different times on the P53 messenger RNA expression in Michigan Cancer Foundation-7 cell. Relative expression levels were obtained using the comparative Ct (ΔΔCt) method (*P < 0.05)
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Results of real-time quantitative polymerase chain reaction before and after thymoquinone treatment at different times on the P53 messenger RNA expression in Michigan Cancer Foundation-7 cell. Relative expression levels were obtained using the comparative Ct (ΔΔCt) method (*P < 0.05)
Mentions: It was suggested that apoptotic induction in cancer cells by TQ requires the activation of P53 gene expression. To examine this hypothesis, we used MCF-7 cell line, as cancerous cell line. We examined the inhibitory effects of 25 μmol/L TQ (based on IC50 index) at different times on the mRNA expression of P53 in MCF-7 cell line. The P53 gene expression was dramatically up-regulated by ascending time, in particular, at 72 h treatment its expression was increased significantly [Figure 3, P < 0.05]. Using Dunnett test, average of P53 gene expression at different times was 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