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A ‘turn-on’ fluorescent probe for lysosomal phosphatase: a comparative study for labeling of cancer cells†
Journal of Materials Chemistry B ( IF 7 ) Pub Date : 2018-06-26 00:00:00 , DOI: 10.1039/c8tb01143e
Arup Podder 1, 2, 3, 4, 5 , Sudipta Senapati 5, 6, 7, 8 , Pralay Maiti 5, 6, 7, 8 , Devaraj Kamalraj 4, 5, 9, 10 , Syed S Jaffer 4, 5, 9, 10 , Sabina Khatun 1, 2, 3, 4, 5 , Sankarprasad Bhuniya 1, 2, 3, 4, 5
Affiliation  

We have described the ability of a newly synthesized fluorescent probe (LP1) to detect phosphatase activity in lysosomes in cancer cells. Probe LP1 displayed a 33-fold fluorescence intensity enhancement at λem 532 nm in the presence of phosphatase in HEPES buffer (pH 4.5). The quantum yield of probe LP1 was increased by ∼21-fold upon exposure to phosphatase at acidic pH. The probe LP1 is highly chemoselective toward phosphatase (ALP/ACP) and is insensitive to interference by ubiquitous biological analytes. The high cell adhesion property and cell viability of LP1 indicate that LP1 is biocompatible and nontoxic; these two characteristic features make it a suitable candidate for phosphatase tracking in living cells. LP1 dose-dependent fluorescence images in living cells suggested that the expression of phosphatase in cancer cells (HeLa) is 2-fold higher as compared to the normal NIH-3T3 cells. The colocalization images confirmed that LP1 was exclusively localized in lysosomes. We envision that LP1 could be a potential tool in clinical diagnosis for discriminating cancer cells from normal cells depending on the expression of phosphatase in lysosomes.

中文翻译:

溶酶体磷酸酶的“开启”荧光探针:癌细胞标记的比较研究

我们已经描述了新合成的荧光探针(LP1)检测癌细胞中溶酶体中磷酸酶活性的能力。探针LP1显示的33倍的荧光强度增强,在λ EM在磷酸酶在HEPES缓冲液(pH 4.5)的存在532纳米。在酸性pH下暴露于磷酸酶后,探针LP1的量子产率提高了约21倍。探针LP1对磷酸酶(ALP / ACP)具有高度的化学选择性,并且对普遍存在的生物分析物的干扰不敏感。LP1的高细胞粘附特性和细胞生存能力表明LP1具有生物相容性且无毒;这两个特征使其成为活细胞中磷酸酶追踪的合适候选者。活细胞中LP1剂量依赖性荧光图像表明,癌细胞(HeLa)中的磷酸酶表达比正常NIH-3T3细胞高2倍。共定位图像证实LP1仅位于溶酶体中。我们设想,LP1可能是临床诊断中用于区分癌细胞与正常细胞的潜在工具,具体取决于溶酶体中磷酸酶的表达。
更新日期:2018-06-26
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