Original article
General thoracic
Effectiveness of Trichostatin A as a Potential Candidate for Anticancer Therapy in Non–Small-Cell Lung Cancer

https://doi.org/10.1016/j.athoracsur.2005.06.059Get rights and content

Background

A well-known histone deacetylase inhibitor, trichostatin A, was applied to non–small-cell lung cancer cells to determine whether inhibition of histone deacetylase leads to the production of proteins that either arrest tumor cell growth or lead to tumor cell death.

Methods

Trichostatin A (0.01 to 1.0 μmol/L) was applied to one normal lung fibroblast and four non–small-cell lung cancer lines, and its effect was determined by flow cytometry, annexin-V staining, immunoprecipitation, and Western blot analysis.

Results

Trichostatin A demonstrated tenfold greater growth inhibition in all four non–small-cell lung cancer lines compared with normal controls, with a concentration producing 50% inhibition ranging from 0.01 to 0.04 μmol/L for the tumor cell lines and 0.7 μmol/L for the normal lung fibroblast line. Trichostatin A treatment reduced the percentage of cells in S phase (10% to 23%) and increased G1 populations (10% to 40%) as determined by flow cytometry. Both annexin-V binding assay and upregulation of the protein, gelsolin (threefold to tenfold), demonstrated that the tumor cells were apoptotic, whereas normal cells were predominantly in cell cycle arrest. Trichostatin A increased histone H4 acetylation and expression of p21 twofold to 15-fold without significant effect on p16, p27, CDK2, and cyclin D1.

Conclusions

Collectively, these data suggest that inhibition of histone deacetylation may provide a valuable approach for lung cancer treatment. We evaluated trichostatin A as a potential candidate for anticancer therapy in non–small-cell lung cancer.

Section snippets

Reagents

Dulbecco’s modified Eagle’s medium, RPMI 1640 medium, fetal bovine serum, trypsin, phosphate-buffered saline, penicillin-streptomycin, and glutamate were purchased from Invitrogen Life Technologies (Carlsbad, CA). Trichostatin A was purchased from Calbiochem (San Diego, CA). Bovine serum albumin and soybean trypsin inhibitor were from Sigma Chemical Company (St. Louis, MO). Proto-gel was from National Diagnostic (Atlanta, GA). Anti-acetylated histone H4 was from Upstate Biotechnology (Lake

Trichostatin A Possesses Antiproliferative Activity Against Non–Small-Cell Lung Cancer Cells

Proliferation in all four NSCLC cell lines was inhibited by TSA in a dose-dependent fashion (Table 1). We determined that effective growth inhibition by TSA occurred within 48 hours. After 48 hours of incubation, the IC50 of the two squamous lines (Calu-1 and H520) was 0.034 and 0.046 μmol/L, respectively. The IC50 of the two adenocarcinoma lines (H441 and H23) was 0.012 and 0.054 μmol/L, respectively. The mean IC50 for all four NSCLC lines was 0.036 ± 0.019 μmol/L. In contrast to the tumor

Comment

The antiproliferative properties of TSA were characterized in four NSCLC lines and in one NLF line. Differential effects in normal versus tumor cells were observed because certain concentrations of TSA produced significant cell death in NSCLC cells compared with normal controls, suggesting the possibility of a therapeutic window. The IC50, as determined on cell growth inhibition assay, is well below the physiologic dose. However, the significance of our observation needs to be further evaluated

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