Dastjerdi Mehdi Nikbakht, Zamani Saeed, Mardani Mohammad, Beni Batool Hashemi
Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.
Res Pharm Sci. 2016 Dec;11(6):505-512. doi: 10.4103/1735-5362.194899.
In this study, we evaluated the effects of all-trans retinoic acid (ATRA) alone or in combination with genistein (GEN) in p14 tumor suppressor gene and subsequent apoptosis of human ovarian carcinoma cells (OVCAR-3). The cells were treated with ATRA or GEN at concentrations of 50 and 25 μM respectively, either alone or in combination for 24 and 48 h. The cell viability was evaluated using 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. The percentage of cell apoptosis was determined using flow cytometry and p14 gene expression was measured using real time PCR. The MTT results showed that in both ATRA and GEN treated groups, the cell viabilityviability in group treated for 48 h was significantly lower than group treated for 24 h. The flow cytometry results showed that the percentage of apoptotic cells in groups that treated with ATRA and GEN in combination for 24 h and 48 h was significantly more than all other tested groups. The real time results showed that the mRNA level of p14 in cells treated with both drugs for 48 h was significantly higher than all other groups. In conclusion, we confirm that GEN in combination with ATRA is an effective strategy to up regulate the p14 tumor suppressor gene and induce cell apoptosis in OVCAR-3 cell line.
在本研究中,我们评估了全反式维甲酸(ATRA)单独使用或与染料木黄酮(GEN)联合使用对p14肿瘤抑制基因以及人卵巢癌细胞(OVCAR-3)后续凋亡的影响。细胞分别用浓度为50 μM的ATRA或25 μM的GEN处理,单独处理或联合处理24小时和48小时。使用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法评估细胞活力。使用流式细胞术测定细胞凋亡百分比,并使用实时PCR测量p14基因表达。MTT结果表明,在ATRA和GEN处理组中,处理48小时组的细胞活力明显低于处理24小时组。流式细胞术结果表明,联合使用ATRA和GEN处理24小时和48小时组的凋亡细胞百分比明显高于所有其他测试组。实时结果表明,两种药物处理48小时的细胞中p14的mRNA水平明显高于所有其他组。总之,我们证实GEN与ATRA联合使用是上调p14肿瘤抑制基因并诱导OVCAR-3细胞系细胞凋亡的有效策略。