The impact of different extracellular matrices on melatonin effect in proliferation and stemness properties of ovarian cancer cells
The impact of different extracellular matrices on melatonin effect in proliferation and stemness properties of ovarian cancer cells
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DOI：
10.1016/j.biopha.2016.12.119
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发表时间：
2017-03-01
影响因子：
7.5
通讯作者：
Nouri, Mohammad
中科院分区：
文献类型：
--
作者：
Akbarzadeh, Maryam;Rahbarghazi, Reza;Nouri, Mohammad
目的：内源性褪黑素在调节不同器官功能方面具有多种生理作用。最近的研究揭示了该分子对肿瘤发展的抑瘤和保护作用。在这项研究中，我们研究了褪黑激素对不同类型细胞外基质中SKOV3卵巢癌细胞的干性、形态、侵袭性和活力的影响及其关键机制。方法：采用MTT法测定细胞活力。将4 × 10(3)个细胞接种于6个孔板上的不同基质上，进行菌落形成试验。应用针对干细胞标记物CD133和CD44的抗体后，通过流式细胞术测定肿瘤干细胞样细胞的百分比。不同类型的细胞外基质，包括纤维连接蛋白、明胶、胶原蛋白和基质，在褪黑激素存在的情况下培养细胞。通过Real-time PCR检测下游基因VEGF、E-cadherin的表达。结果：褪黑素（0.1 mM）可使活细胞率降低61.79±8.2% （p < 0.05）。集落形成实验显示褪黑素对这些细胞的集落形成有显著的抑制作用。褪黑素对明胶的影响最大（p < 0.05）。干细胞标记物鉴定表明，与其他类型的细胞外基质相比，使用matrigel可显著增加肿瘤干细胞样细胞的百分比（p < 0.05）。然而，褪黑素略微减少了所有培养基质中干细胞样细胞的数量。我们的基因表达分析结果显示，褪黑素诱导E-cadherin显著增加，VEGF表达水平显著降低（p < 0.05）。结论：卵巢癌细胞与邻近基质的相互作用决定了褪黑素的抗侵袭活性。(C) 2016 Elsevier Masson SAS。版权所有。
Aim: Endogenous melatonin has numerous physiological roles on modulating the function of different organs. Recent studies revealed oncostatic and protective effects of this molecule on tumor development. In this study, we examined the impact of melatonin and key underlying mechanisms on stemness, morphology, invasiveness and viability of SKOV3 ovarian cancer cells in different types of extracellular matrix.Methods: Cell viability was evaluated by MTT Assay. Colony-forming assay was performed by seeding 4 x 10(3) cells on different matrices in six well-plate. The percentage of cancer stem like cells was determined by flow cytometric assay after applying antibodies against stemness markers, CD133 and CD44. Different types of extracellular matrix including fibronectin, gelatin, collagen and matrigel were applied to incubate the cells in the presence of melatonin. Downstream gene expressions including VEGF and E-cadherin were determined by Real-time PCR.Results: Melatonin (0.1 mM) decreased the percentage of viable cells up to 61.79 +/- 8.2% (p < 0.05). Colony formation assay revealed the significant impact of melatonin in inhibition of colony formation in these cells. The maximum effect was shown in the cells incubated with melatonin on gelatin (p < 0.05). Identification of stemness markers showed that applying matrigel caused significant increase in the percentage of cancer stem like cells compared to other types of extracellular matrix (p < 0.05). However melatonin slightly diminished the number of stem cell like cells in all incubated matrices. Our results from gene expression assays revealed that melatonin induced a marked increase in E-cadherin along with decrease in VEGF expression levels (p < 0.05).Conclusion: Our results suggest that interaction between ovarian cancer cells and neighboring matrices determines the subsequent anti invasive activities of melatonin. (C) 2016 Elsevier Masson SAS. All rights reserved.