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使用毛细管电泳非接触电导检测(CE-C D)研究 Tamoxifen 耐药 MCF-7 细胞中乳酸和丙酮酸水平与乳酸脱氢酶基因表达和葡萄糖消耗的相关性。

Lactate and pyruvate levels correlation with lactate dehydrogenase gene expression and glucose consumption in Tamoxifen-resistant MCF-7 cells using capillary electrophoresis with contactless conductivity detection (CE-C D).

机构信息

Department of Pharmacy, Faculty of Pharmacy, Al-Zaytoonah University of Jordan, Amman, Jordan.

Department of Analytical Chemistry, Faculty of Pharmacy, Helwan University, Cairo, Egypt.

出版信息

Electrophoresis. 2022 Feb;43(3):446-455. doi: 10.1002/elps.202100217. Epub 2021 Nov 1.

Abstract

Breast cancer is the second leading cause of cancer death in women after lung cancer. The first-line treatment of metastatic breast cancer in premenopausal women relies on tamoxifen. The development of tamoxifen resistance is not fully understood. In this study, capillary electrophoresis with capacitively coupled contactless conductivity detector was developed to monitor the changes in lactate and pyruvate levels in supernatant media of three models of developed MCF-7 tamoxifen-resistant cells and correlate these metabolites changes with lactate dehydrogenase genes expression and glucose consumption. The electrophoretic separation was achieved under reversed electroosmotic flow conditions. The linear ranges were 0.15-5 and 0.01-1 mM with a correlation coefficient of 0.9966 and 0.9971 and the limits of detection were 0.01 and 0.02 μM for lactate and pyruvate, respectively. Inter- and intrarun accuracy were in the range of 96.88-105.94% with precision (CV, %) of ≤7.35%. The method was completely validated and the results were in agreement with those obtained using the lactate and glucose assay kits. The results revealed a significant increase in both lactate and pyruvate production in the three tamoxifen-resistant MCF-7 cells models compared to control cells. This increase was correlated with the increase of lactate dehydrogenase genes expression and the increase of glucose consumption.

摘要

乳腺癌是女性癌症死亡的第二大主要原因,仅次于肺癌。绝经前转移性乳腺癌的一线治疗依赖于他莫昔芬。他莫昔芬耐药的发展机制尚未完全阐明。在这项研究中,我们开发了一种毛细管电泳-电容耦合非接触式电导检测器,用于监测三种已建立的 MCF-7 他莫昔芬耐药细胞模型上清液中乳酸和丙酮酸水平的变化,并将这些代谢物的变化与乳酸脱氢酶基因表达和葡萄糖消耗相关联。在反电渗流条件下实现电泳分离。乳酸和丙酮酸的线性范围分别为 0.15-5 和 0.01-1 mM,相关系数分别为 0.9966 和 0.9971,检测限分别为 0.01 和 0.02 μM。日内和日间准确度在 96.88-105.94%范围内,精密度(CV,%)≤7.35%。该方法已完全验证,结果与使用乳酸和葡萄糖检测试剂盒获得的结果一致。结果表明,与对照细胞相比,三种他莫昔芬耐药 MCF-7 细胞模型中乳酸和丙酮酸的生成均显著增加。这种增加与乳酸脱氢酶基因表达的增加和葡萄糖消耗的增加有关。

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