Dastjerdi Mehdi Nikbakht, Mehdiabady Ebrahim Momeni, Iranpour Farhad Golshan, Bahramian Hamid
Department of Anatomical Sciences and Molecular Biology, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran.
Int J Prev Med. 2016 Apr 14;7:66. doi: 10.4103/2008-7802.180412. eCollection 2016.
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).
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.
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.
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.
多年来,黑种草在医学上一直是一种营养调味成分和多种疾病的天然治疗方法。早期研究报道,其种子中的活性化合物百里醌(TQ)具有抗癌特性。先前的研究表明,TQ可诱导乳腺癌细胞凋亡,但P53在凋亡途径中的作用尚不清楚。因此,本研究报道了TQ对肿瘤抑制基因P53表达的影响以及对乳腺癌细胞系密歇根癌症基金会-7(MCF-7)凋亡诱导的作用。
培养MCF-7细胞系并用TQ处理,处理24小时后进行3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)测定以评估半数抑制浓度(IC50)值。通过流式细胞术测量凋亡细胞的百分比。进行实时聚合酶链反应(PCR)以估计不同时间MCF-7细胞系中P53的信使核糖核酸表达。
使用MTT测定法确定TQ在MCF-7细胞中的IC50值为25μM。流式细胞术和实时PCR结果表明,TQ可诱导MCF-7细胞凋亡,且P53基因表达分别随时间显著上调。因此,在48小时和72小时时有显著差异。
我们的结果表明,TQ可通过时间依赖性方式上调乳腺癌细胞系(MCF-7)中P53的表达来诱导MCF-7细胞凋亡。