Premium
High‐throughput Screening for Identification of Inhibitors of Ep CAM ‐Dependent Growth of Hepatocellular Carcinoma Cells
Author(s) -
Henrich Curtis J.,
Budhu Anuradha,
Yu Zhipeng,
Evans Jason R.,
Goncharova Ekaterina I.,
Ransom Tanya T.,
Wang Xin W.,
McMahon James B.
Publication year - 2013
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/cbdd.12146
Subject(s) - hepatocellular carcinoma , high throughput screening , identification (biology) , throughput , cancer research , chemistry , virtual screening , computational biology , biochemistry , biology , drug discovery , computer science , botany , telecommunications , wireless
The cancer stem cell marker, Ep CAM , is an important indicator of Wnt/β‐catenin signaling activation and a functional component of hepatocellular tumor‐initiating cells. A high‐throughput screening assay was developed to identify inhibitors of Ep CAM ‐dependent growth of hepatocellular carcinoma (HCC) cells. Ep CAM (+) and Ep CAM (−) HCC cell lines were assessed for differential sensitivity to a Wnt/β‐catenin pathway inhibitor. Libraries comprising 22 668 pure compounds and 107 741 crude or partially purified natural product extracts were tested, and 12 pure compounds and 67 natural product extracts were identified for further study. Three active compounds and the positive control were further characterized in terms of effects on Ep CAM expression. Treatment of Ep CAM (+) Hep3B cells resulted in loss of Ep CAM expression as assessed by flow cytometry. This reduction was incomplete (most cells continued to express Ep CAM ), but resulted in generation of cell populations expressing lower levels of Ep CAM . Sublethal concentrations (~ IC 50 ) reduced median Ep CAM expression to 28% of control after 1 day and 19% of control after 2 days. Reduction in Ep CAM expression preceded growth inhibition suggesting that a threshold of Ep CAM expression may be required for growth of Ep CAM ‐dependent cells. The identification of compounds with a variety of possible molecular targets suggests a likelihood of multiple mechanisms for modulation of Ep CAM ‐dependent cell growth.