Etemad-Moghadam S, Fouladdel S, Azizi E, Alaeddini M
Dental Research Center, Tehran University of Medical Sciences, Tehran, Iran.
J BUON. 2013 Oct-Dec;18(4):1062-8.
Aberrant proliferation is an essential feature of cancer cells, which can be caused by alterations in components of the cell cycle, such as minichromosome maintenance protein-3 (MCM3) and Ki-67. Doxorubicin is a cytotoxic/cytostatic anticancer agent commonly used in chemotherapy. We investigated the effect of this drug on MCM3 and Ki-67 in the KB cell line, which is considered a subline of HeLa cell line.
KB cells were treated with doxorubicin and its effect on apoptosis, mRNA levels and protein expression of MCM3 and Ki-67 was determined by flow cytometry (annexin V-FITC/PI assay), quantitative real-time RT-PCR (qRT-PCR) and immunocytochemistry, respectively. Cytotoxicity was assessed using the MTT assay. One-way analysis of variance (ANOVA) was used for comparing groups and differences were assessed by a Tukey's post hoc test.
Protein expression of both biomarkers and MCM3 mRNA were not affected by doxorubicin, but Ki-67 mRNA significantly increased after treatment (p=0.049).
Considering that doxorubicin can influence certain biochemical events that lead to modifications in Ki- 67, this factor might be useful in evaluating the impact of anthracycline-based chemotherapeutic agents. Changes in MCM3 following doxorubicin treatment require further investigation.
异常增殖是癌细胞的一个基本特征，其可由细胞周期组分的改变引起，如微小染色体维持蛋白3（MCM3）和Ki-67。阿霉素是一种常用于化疗的细胞毒性/细胞生长抑制性抗癌药物。我们研究了该药物对KB细胞系中MCM3和Ki-67的影响，KB细胞系被认为是HeLa细胞系的一个亚系。
用阿霉素处理KB细胞，分别通过流式细胞术（膜联蛋白V-异硫氰酸荧光素/碘化丙啶检测法）、定量实时逆转录聚合酶链反应（qRT-PCR）和免疫细胞化学法测定其对细胞凋亡、MCM3和Ki-67的mRNA水平及蛋白表达的影响。使用MTT检测法评估细胞毒性。采用单因素方差分析（ANOVA）比较各组，并通过Tukey事后检验评估差异。
两种生物标志物的蛋白表达及MCM3 mRNA均不受阿霉素影响，但处理后Ki-67 mRNA显著增加（p = 0.049）。
鉴于阿霉素可影响某些导致Ki-67发生改变的生化事件，该因子可能有助于评估基于蒽环类的化疗药物的影响。阿霉素处理后MCM3的变化需要进一步研究。
