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. 2019 Dec 6;11(12):1964. doi: 10.3390/cancers11121964

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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HCPT decreases TOP I enzyme activity and inhibits its expression in ESCC. (A) Cell lysates were analyzed after HCPT treatment with the indicated concentration for 24 h. TOP I enzyme was extracted following the producer’s protocol (www.topogen.com). The activity of TOP I was measured and visualized by DNA electrophoresis as the indicated amount of supercoiled DNA. (B) ESCC cells were harvested and lysated with HCPT treatment for 48 and 72 h and the expression of TOP I was visualized by Western blot method. β-actin was used as an internal reference control. (C) Phosphorylation of rH2A.X at serine 139 site indicated DNA foci formation and damage, which is the late-stage phenomenon after TOP I activity was broken down. Western blot was carried out for developing the expression of H2A.XS139 after HCPT treatment at 48 h in the indicated concentration. β-actin was used as an internal reference control. Three independent repeats were performed for each experiment.