title: A rapid fluorescent screening method for cellular sensitivity to anti-cancer compound.
abstract: The tetrazolium salts (MTT, XTT, MTS, WST) based colorimetric assay or resazurin based fluorimetric assay are currently typical methods for cell sensitivity determination to anticancer compounds. We presented here a new rapid method for this purpose. This method uses a fluorescent dye named DCFH-DA which is previously taken as a intracellular probe for measurement of H(2)O(2) levels within a cell. The application basis for this method lies in two facts: the membrane permeable feature of the final metabolite of DCFH-DA inside a cell, and the linearity relationship between cell number and H(2)O(2) level. The results showed that there was a perfect association between cell number and fluorescent intensity determined by the DCFH-DA method, no matter whether using resuspended or adherent cells, and further 50% concentration of inhibition (IC(50)) comparison between data obtained by DCFH-DA method or MTT method using a positive known anticancer compound Baicalin showed that there were no significant differences in cellular sensitivity determination to compound Baicalin though there existed a relatively higher coefficient of variation of IC(50) by the DCFH-DA method than that by the MTT method. Thus our data indicate that DCFH-DA might not only be a fine reagent for determination of H(2)O(2) levels in cells but also an ideal fluorescent dye for cellular sensitivity test of anti-cancer compounds, and may be suitable for primary high-throughput drugs screening.
authors.affiliation: Lab of Microbial and Biochemical Pharmaceutical, School of Pharmacy, Shanghai Jiaotong University, 800 Dong Chuan Road, Minhang District, Shanghai, 200240, China.
authors.forename: Wenwei
authors.initials: W
authors.lastname: Mao
authors.forename: Xinlin
authors.initials: X
authors.lastname: Chen
authors.forename: Tian
authors.initials: T
authors.lastname: Yang
authors.forename: Yu
authors.initials: Y
authors.lastname: Yin
authors.forename: Mei
authors.initials: M
authors.lastname: Ge
authors.forename: Minyu
authors.initials: M
authors.lastname: Luo
authors.forename: Daijie
authors.initials: D
authors.lastname: Chen
authors.forename: Xiuping
authors.initials: X
authors.lastname: Qian