Department of Obsterics and Gynaecology in Ruda Slaska, Medical University of Silesia, Ruda Slaska, Poland
Department of Gynecology and Obstetrics with Gynecologic Oncology, Ludwik Rydygier Memorial Specialized Hospital, Kraków, Poland
Curr Pharm Biotechnol. 2020;21(12):1269-1277. doi: 10.2174/1389201021666200513074022.
Salinomycin is part of a group of ionophore antibiotics characterized by an activity towards tumor cells. To this day, the mechanism through which salinomycin induces their apoptosis is not fully known yet. The goal of this study was to assess the expression pattern of genes and the proteins coded by them connected with the process of programmed cell death in an endometrial cancer cell Ishikawa culture exposed to salinomycin and compared to the control.
Analysis of the effect of salinomycin on Ishikawa endometrial cancer cells (ECACC 99040201) included a cytotoxicity MTT test (with a concentration range of 0.1-100 μM), assessment of the induction of apoptosis and necrosis by salinomycin at a concentration of 1 μM as well the assessment of the expression of the genes chosen in the microarray experiment (microarray HG-U 133A_2) and the proteins coded by them connected with apoptosis (RTqPCR, ELISA assay). The statistical significance level for all analyses carried out as part of this study was p<0.05.
It was observed that salinomycin causes the death of about 50% of cells treated by it (50.74±0.80% of all cells) at a concentration of 1μM. The decrease in the number of living cells was determined directly after treatment of the cells with the drug (time 0). The average percent of late apoptotic cells was 1.65±0.24% and 0.57±0.01% for necrotic cells throughout the entire observation period.
Microarray analysis indicated the following number of mRNA differentiating the culture depending on the time of incubation with the drug: H_12 vs C = 114 mRNA, H_8 vs C = 84 mRNA, H_48 vs. C = 27 mRNA, whereas 5 mRNAs were expressed differently at all times. During the whole incubation period of the cells with the drug, the following dependence of the expression profile of the analyzed transcripts was observed: Bax>p53>FASL>BIRC5>BCL2L.
The analysis carried out indicated that salinomycin, at a concentration of 1 μM, stopped the proliferation of 50% of endometrial cancer cells, mainly by inducing the apoptotic process of the cells. The molecular exponent of the induction of programmed cell death was an observed increase in the transcriptional activity of pro-apoptotic genes: Bax;p53;FASL and a decrease in the expression of anti-apoptotic genes: BCL2L2; BIRC5.
盐霉素是一类具有抗肿瘤细胞活性的离子载体抗生素的一部分。迄今为止，盐霉素诱导其凋亡的机制尚未完全阐明。本研究的目的是评估暴露于盐霉素的子宫内膜癌细胞 Ishikawa 培养物中与细胞程序性死亡过程相关的基因及其编码蛋白的表达模式，并与对照组进行比较。
分析盐霉素对 Ishikawa 子宫内膜癌细胞（ECACC 99040201）的影响包括细胞毒性 MTT 试验（浓度范围为 0.1-100 μM），评估 1 μM 盐霉素诱导的细胞凋亡和坏死，以及选择的基因的表达微阵列实验（微阵列 HG-U 133A_2）及其与凋亡相关的编码蛋白（RTqPCR、ELISA 测定）。本研究中进行的所有分析的统计学显著性水平均为 p<0.05。
结果表明，盐霉素在 1μM 浓度下可导致约 50%的处理细胞死亡（所有细胞的 50.74±0.80%）。在直接用药物处理细胞后，活细胞数量减少（时间 0）。在整个观察期间，晚期凋亡细胞的平均百分比为 1.65±0.24%，坏死细胞的平均百分比为 0.57±0.01%。
微阵列分析表明，根据与药物孵育时间的不同，培养物中存在以下数量的 mRNA 差异：H_12 vs C = 114 mRNA，H_8 vs C = 84 mRNA，H_48 vs C = 27 mRNA，而在所有时间点都有 5 种 mRNA 表达不同。在整个细胞与药物孵育期间，观察到以下分析转录本表达谱的依赖性：Bax>p53>FASL>BIRC5>BCL2L。
分析表明，盐霉素在 1 μM 浓度下可阻止 50%的子宫内膜癌细胞增殖，主要通过诱导细胞凋亡过程。程序性细胞死亡的分子指数是观察到的促凋亡基因转录活性的增加：Bax;p53;FASL 和抗凋亡基因表达的减少：BCL2L2;BIRC5。
