T Kozak, O Lykhova, T Serhiichuk, N Bezdieniezhnykh, V Chekhun
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引用次数: 0
Abstract
Background: The development of human breast cancer (BC) is known to be closely related to disturbances in the mammary gland microbiota. Bacteria of the genus Bifidobacterium are an important component of normal breast microbiota and exert antitumor activity. The molecular-biological mechanisms of interaction between BC cells and microbiota members remain poorly studied yet. The aim of this study was to develop and optimize an experimental model system for the co-cultivation of BC cells with Bifidobacterium animalis in vitro.
Materials and methods: Human ВС cells of the MCF-7, T47D, and MDA-MB-231 lines, as well as live and heat-inactivated bacteria of Bifidobacterium animalis subsp. lactis (B. animalis) were used as research objects. The growth kinetics and viability of B. animalis in the presence of different ВС cell lines and without them were determined by both the turbidimetry method and seeding on an elective nutrient medium. Glucose consumption and lactate production by bifidobacteria were assessed by biochemical methods. The viability of BC cells was determined by a standard colorimetric method.
Results: The growth kinetics of B. animalis in the complete DMEM nutrient medium showed standard patterns. The indicators of glucose consumption and lactate production of B. animalis confirm its physiological metabolic activity under the growth conditions. The presence of BC cells in the model system did not affect the duration of the growth phases of the B. animalis cells' population but contributed to the increase in their counts. A significant decrease in the number of live BC cells of all studied lines was observed only after 48 h of co-cultivation with live B. animalis. To achieve similar suppression of the BC cell viability, 10-30-fold higher counts of heatinactivated bacteria were required compared to live ones.
Conclusions: The optimal conditions for co-cultivation of human BC cells and living B. animalis cells in vitro have been identified.
背景：众所周知，人类乳腺癌（BC）的发生与乳腺微生物群的紊乱密切相关。双歧杆菌属细菌是正常乳腺微生物群的重要组成部分，具有抗肿瘤活性。有关 BC 细胞与微生物群成员之间相互作用的分子生物学机制的研究仍然很少。材料与方法：以 MCF-7、T47D 和 MDA-MB-231 株系的人类ВС细胞以及动物双歧杆菌亚种（Bifidobacterium animalis subsp.lactis，B. animalis）的活菌和热灭活菌为研究对象。通过浊度法和在选择性营养培养基上播种两种方法测定了动物双歧杆菌在有不同ВС细胞系存在和无ВС细胞系存在时的生长动力学和存活率。双歧杆菌消耗葡萄糖和产生乳酸的情况通过生化方法进行评估。用标准比色法测定双歧杆菌细胞的活力：结果：动物双歧杆菌在完全 DMEM 营养培养基中的生长动力学表现出标准模式。动物芽孢杆菌消耗葡萄糖和产生乳酸的指标证实了其在生长条件下的生理代谢活动。模型系统中 BC 细胞的存在并不影响 B. animalis 细胞群生长阶段的持续时间，但有助于其数量的增加。只有在与活的 B. animalis 共同培养 48 小时后，才能观察到所有研究品系的活 BC 细胞数量明显减少。要达到类似的抑制BC细胞活力的效果，热灭活菌的数量需要比活菌高10-30倍：结论：人类巴氏杆菌细胞与动物巴氏杆菌活细胞体外共培养的最佳条件已经确定。
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