Genetic and small molecule inhibition of arylamine N-acetyltransferase 1 reduces anchorage-independent growth in human breast cancer cell line MDA-MB-231.

Abstract

Arylamine N-acetyltransferase 1 (NAT1) expression is reported to affect proliferation, invasiveness, and growth of cancer cells. MDA-MB-231 breast cancer cells were engineered such that NAT1 expression was elevated or suppressed, or treated with a small molecule inhibitor of NAT1. The MDA-MB-231 human breast cancer cell lines were engineered with a scrambled shRNA, a NAT1 specific shRNA or a NAT1 overexpression cassette stably integrated into a single flippase recognition target (FRT) site facilitating incorporation of these different genetic elements into the same genomic location. NAT1-specific shRNA reduced NAT1 activity in vitro by 39%, increased endogenous acetyl coenzyme A levels by 35%, and reduced anchorage-independent growth (sevenfold) without significant effects on cell morphology, growth rates, anchorage-dependent colony formation, or invasiveness compared to the scrambled shRNA cell line. Despite 12-fold overexpression of NAT1 activity in the NAT1 overexpression cassette transfected MDA-MB-231 cell line, doubling time, anchorage-dependent cell growth, anchorage-independent cell growth, and relative invasiveness were not changed significantly when compared to the scrambled shRNA cell line. A small molecule (5E)-[5-(4-hydroxy-3,5-diiodobenzylidene)-2-thioxo-1,3-thiazolidin-4-one (5-HDST) was 25-fold more selective towards the inhibition of recombinant human NAT1 than N-acetyltransferase 2. Incubation of MDA-MB-231 cell line with 5-HDST resulted in 60% reduction in NAT1 activity and significant decreases in cell growth, anchorage-dependent growth, and anchorage-independent growth. In summary, inhibition of NAT1 activity by either shRNA or 5-HDST reduced anchorage-independent growth in the MDA-MB-231 human breast cancer cell line. These findings suggest that human NAT1 could serve as a target for the prevention and/or treatment of breast cancer.

• Authors: Marcus W Stepp, Mark A Doll, Samantha M Carlisle, J Christopher States, David W Hein
• Journal: Molecular carcinogenesis (Mol Carcinog) Vol. 57 Issue 4 Pg. 549-558 (04 2018) ISSN: 1098-2744 [Electronic] United States
• PMID: 29315819 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: © 2018 Wiley Periodicals, Inc.

Chemical References

• Isoenzymes
• RNA, Small Interfering
• Small Molecule Libraries
• Thiazolidines
• Arylamine N-Acetyltransferase
• N-acetyltransferase 1

Topics

• Arylamine N-Acetyltransferase (antagonists & inhibitors, genetics, metabolism)
• Binding Sites (genetics)
• Breast Neoplasms (enzymology, genetics, pathology)
• Cell Adhesion (drug effects, genetics)
• Cell Line, Tumor
• Cell Proliferation (drug effects, genetics)
• Female
• Gene Expression Regulation, Enzymologic (drug effects)
• Gene Expression Regulation, Neoplastic (drug effects)
• Humans
• Isoenzymes (antagonists & inhibitors, genetics, metabolism)
• Molecular Structure
• RNA Interference
• RNA, Small Interfering (genetics)
• Small Molecule Libraries (chemistry, pharmacology)
• Thiazolidines (chemistry, pharmacology)