title: Development of an In Vitro Model to Screen CYP1B1-Targeted Anticancer Prodrugs.
abstract: Cytochrome P450 1B1 (CYP1B1) is an anticancer therapeutic target due to its overexpression in a number of steroid hormone-related cancers. One anticancer drug discovery strategy is to develop prodrugs specifically activated by CYP1B1 in malignant tissues to cytotoxic metabolites. Here, we aimed to develop an in vitro screening model for CYP1B1-targeted anticancer prodrugs using the KLE human endometrial carcinoma cell line. KLE cells demonstrated superior stability of CYP1B1 expression relative to transiently transfected cells and did not express any appreciable amount of cognate CYP1A1 or CYP1A2, which would have compromised the specificity of the screening assay. The effect of two CYP1B1-targeted probe prodrugs on KLE cells was evaluated in the absence and presence of a CYP1B1 inhibitor to chemically "knock out" CYP1B1 activity (CYP1B1 inhibited). Both probe prodrugs were more toxic to KLE cells than to CYP1B1-inhibited KLE cells and significantly induced G0/G1 arrest and decreased the S phase in KLE cells. They also exhibited pro-apoptotic effects in KLE cells, which were attenuated in CYP1B1-inhibited KLE cells. In summary, a KLE cell-based model has been characterized to be suitable for identifying CYP1B1-targeted anticancer prodrugs and should be further developed and employed for screening chemical libraries.
authors.affiliation: 1 School of Pharmacy, Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA.
authors.forename: Zhiying
authors.initials: Z
authors.lastname: Wang
authors.affiliation: 1 School of Pharmacy, Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA.
authors.forename: Yao
authors.initials: Y
authors.lastname: Chen
authors.affiliation: 1 School of Pharmacy, Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA.
authors.forename: Laura M
authors.initials: LM
authors.lastname: Drbohlav
authors.affiliation: 2 School of Pharmacy, Department of Medicinal Chemistry, University of Kansas, Lawrence, KS, USA.
authors.forename: Judy Qiju
authors.initials: JQ
authors.lastname: Wu
authors.affiliation: 1 School of Pharmacy, Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA.
authors.forename: Michael Zhuo
authors.initials: MZ
authors.lastname: Wang
mesh_terms: D000970:Antineoplastic Agents / Q000302:isolation & purification* / Q000494:pharmacology; D045744:Cell Line, Tumor; D065633:Cytochrome P-450 CYP1B1 / Q000037:antagonists & inhibitors* / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D018390:Gene Targeting / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D058990:Molecular Targeted Therapy; D009369:Neoplasms / Q000188:drug therapy*; D011355:Prodrugs / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D011355:Prodrugs; C585496:CYP1B1 protein, human; D065633:Cytochrome P-450 CYP1B1
keywords: CYP1B1; drug screening; endometrial cancer; prodrug; targeted anticancer