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Targeting the Anaphase Promoting Complex to Prevent Lung Cancer Cell Growth
Author(s) -
Wainscott Nathan W,
Hoffman J. Mason,
Taylor B. Frazier,
Trent John O.,
States J. Christopher
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.823.12
Subject(s) - spindle checkpoint , mitosis , anaphase promoting complex , anaphase , microbiology and biotechnology , mitotic catastrophe , metaphase , spindle apparatus , biology , cell growth , a549 cell , cell cycle checkpoint , cancer cell , cell cycle , cancer research , cell , chemistry , cancer , cell division , biochemistry , genetics , gene , chromosome
Background Current chemotherapeutic agents widely used for the treatment of lung cancer include mitotic inhibitors such as taxanes. Taxanes stabilize the mitotic spindle, maintaining the activity of the Spindle Assembly Checkpoint (SAC), thus preventing the metaphase to anaphase transition. Prolonged mitotic arrest results in cell death. Non‐small cell lung cancer (NSCLC) accounts for 80–85% of lung cancer cases. NSCLC may intrinsically be SAC deficient or acquire defects, making them resistant to these drugs. New treatments are needed for SAC deficient tumors. One potential new target for mitotic inhibition is the Anaphase Promoting Complex/Cyclosome (APC/C), an E3 ubiquitin ligase that drives the transition from metaphase to anaphase. The catalytic core of the APC/C consists of the subunits ANAPC2 and ANAPC11. Hypothesis Targeting ANAPC2 to prevent binding of ANAPC11 to ANAPC2 will inhibit the Anaphase Promoting Complex/Cyclosome in lung cancer cells and induce cell death. Methods Four lead compounds (3, 8, 10, and 11) were predicted by in silico analyses to bind the ANAPC2 subunit, and thus inhibit the APC/C by preventing binding of the catalytic subunit, ANAPC11. Compounds 8, 10 and 11 are 2‐amino‐propanol derivatives while compound 3 is a triazaspiro derivative. The compounds were tested for their ability to induce cytotoxicity in two human NSCLC cell lines (H460, A549) and a non‐transformed human bronchial epithelial cell line (HBEC). Cell viability following treatment was evaluated by morphological change by microscopy, Alamar Blue fluorescence assays, and colony forming assays. Results Morphological assessment of cell viability indicated a concentration‐ and time‐dependent increase in the number of mitotic cells with increasing concentration for compounds 3, 8 and 10 in all three cell lines, and evidence of cell death at higher concentrations in A549 and H460 cells. The Alamar Blue assays and colony forming assays confirmed that these compounds lead to a concentration–dependent decrease in A549 and H460 cell viability and proliferation. Compounds 3, 10 and 11 were more effective at inducing cell death than compound 8. Compound 11 was toxic at all concentrations tested in all three cell lines. Conclusions These results indicate that the APC/C could be a viable target for development of novel chemotherapeutic drugs to treat NSCLC. Support or Funding Information ASPET‐Summer Undergraduate Research Fellowship (NCW); KY Lung Cancer Research Program (JCS,); KY Science and Engineering Foundation (JCS); Kosair Charities Pediatric Oncology Research Program (JOT); NIH P30GM106396 (JOT)