School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, Singapore 637457.
Anal Chem. 2010 Jun 15;82(12):5082-7. doi: 10.1021/ac100074n.
Sensitive detection of extracellular lactate concentrations at a single cell level is of importance for studying the metabolic alterations in tumor progression. A unique nanoscale optical fiber lactate sensor was developed to monitor the extracellular lactate concentrations of cancer cells by immobilizing its nanotip with lactate dehydrogenases, which could catalyze lactate conversion to generate NADH for sensitive fluorescence detection. The results demonstrate that the fabricated nanosensor can successfully detect the extracellular lactate concentrations for single HeLa, MCF-7, and human fetal osteoblast (hFOB) cells, showing that the cancer cells have distinctly higher extracellular lactate concentrations than normal cells as that predicted by Warburg effect. The nanosensor was also employed to investigate the effect of a monocarboxylate transporter inhibitor on the lactate efflux from cancer cells. Different lactate efflux inhibition profiles were obtained for HeLa and MCF-7 cell lines. This work demonstrates that the nanosensor has potential for evaluating the effect of metabolic agents on cancer metabolism and survival.
单细胞水平对外界乳酸浓度的灵敏检测对于研究肿瘤进展中的代谢变化非常重要。本研究开发了一种独特的纳米光纤乳酸传感器,通过将纳米尖端固定化乳酸脱氢酶来监测癌细胞的细胞外乳酸浓度,该酶可以催化乳酸转化为生成 NADH 以进行灵敏的荧光检测。结果表明,所制备的纳米传感器可以成功地检测单个 HeLa、MCF-7 和人胎成骨细胞(hFOB)的细胞外乳酸浓度,表明与沃伯格效应预测的一样,癌细胞的细胞外乳酸浓度明显高于正常细胞。该纳米传感器还用于研究单羧酸转运蛋白抑制剂对癌细胞中乳酸外排的影响。对于 HeLa 和 MCF-7 细胞系,获得了不同的乳酸外排抑制谱。这项工作表明,该纳米传感器具有评估代谢剂对癌症代谢和存活的影响的潜力。