EFFECT OF ADHESIVE LLC CELL PRETREATMENT BY OXAMATE ON THE SURVIVAL INDEXES AFTER TRANSITION TO DE-ADHESIVE GROWTH.

Yu Stepanov, D Kolesnik, Yu Yakshibaeva, G Solyanik
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Abstract

Background: The ability to metabolic reprogramming is a distinctive feature of metastatically active tumor cells. A classic example of metabolic reprogramming, characteristic of almost all malignant cells, is aerobic glycolysis. Therefore, inhibition of glycolysis in tumor cells is considered a promising strategy for antitumor therapy.

Aim: To generate Lewis lung carcinoma (LLC) cell subpopulation after pretreatment by a lactate dehydrogenase (LDH) inhibitor - oxamate in adhesive growth conditions, and then to study the metabolism of this subpopulation in the anchorage-independent growth conditions.

Materials and methods: LLC cells were cultured without oxamate or with 17 mM oxamate in the adhesive growth conditions with the following transition to the anchorage-independent growth conditions without oxamate. A distribution of LLC cells by cell cycle phases, apoptosis rate, levels of reactive oxygen species (ROS), E-cadherin, and vimentin were determined by flow cytometry. Glucose consumption and lactate production were determined by spectrophotometry.

Results: 48-h oxamate treatment in adhesive growth conditions resulted in a 30% decrease of the total number of LLC cells compared to the control. In 72 h after the transfer of both oxamate-treated and control cells into the anchorage-independent growth condition without oxamate, the number of viable cells pretreated with oxamate was reduced by 17% (p < 0.05) compared to the control cells. However, the distribution of cells by cell cycle phases did not differ. In cells pre-treated with oxamate, the rate of glucose consumption decreased by 20% (p < 0.05), ROS generation was reduced by 17%, vimentin expression decreased by 10% while the rate of lactate production was the same in oxamate-pretreated and control cells.

Conclusion: The cytostatic effect of oxamate demonstrated in adhesive growth conditions persisted for 72 h in the anchorage-independent growth conditions. The absence of differences in the cell cycle phase distribution and a decrease in the ROS generation may indicate the initial stage of overcoming the cytostatic effect of oxamate after 72 h of culturing LLC cells in anchorage- independent growth conditions.

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草氨酸盐预处理粘附性 llc 细胞对过渡到非粘附性生长后存活指标的影响。
背景:代谢重编程能力是转移性活性肿瘤细胞的一个显著特征。代谢重编程的一个经典例子是有氧糖酵解,这是几乎所有恶性细胞的特征。因此,抑制肿瘤细胞中的糖酵解被认为是一种很有前途的抗肿瘤治疗策略。目的:采用乳酸脱氢酶(LDH)抑制剂-草酸酯在黏附生长条件下预处理Lewis肺癌(LLC)细胞亚群,并研究该亚群在非锚定生长条件下的代谢情况。材料和方法:将LLC细胞在不含草酸盐和17 mM草酸盐的黏附生长条件下培养,然后过渡到不含草酸盐的非锚定生长条件。流式细胞术检测LLC细胞周期分期、凋亡率、活性氧(ROS)、E-cadherin和vimentin水平的分布。用分光光度法测定葡萄糖消耗量和乳酸产量。结果:在黏附生长条件下,草酸酯处理48小时导致LLC细胞总数比对照组减少30%。将经草酸盐处理的细胞和对照细胞转移到不含草酸盐的非锚定生长条件后72 h,经草酸盐预处理的活细胞数量比对照细胞减少了17% (p < 0.05)。然而,不同细胞周期的细胞分布没有差异。在经草酸预处理的细胞中,葡萄糖消耗率降低了20% (p < 0.05), ROS生成减少了17%,vimentin表达降低了10%,而乳酸生成率与对照细胞相同。结论:在不依赖锚定生长条件下,草酸酯在黏附生长条件下的细胞抑制作用可持续72 h。在不依赖锚定生长条件下培养LLC细胞72 h后,细胞周期期分布没有差异,ROS生成减少,这可能表明LLC细胞在克服草酸酯的细胞抑制作用的初始阶段。
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