Molecular determinants of the antitumor effects of trichostatin A in pancreatic cancer cells

作者：Emonds Elisabeth; Fitzner Brit; Jaster Robert^*

来源：World Journal of Gastroenterology, 2010, 16(16): 1970-1978.

DOI：10.3748/wjg.v16.116.1970

• Pancreatic cancer
• Histone deacetylase inhibitor
• Trichostatin-A
• KRAS
• MAP kinases
• p21(Waf1)
• AKT

AIM: To gain molecular insights into the action of the histone deacetylase inhibitor (HDACI) trichostatin-A (TSA) in pancreatic cancer (PC) cells.
METHODS: Three PC cell lines, BxPC-3, AsPC-1 and CAPAN-1, were treated with various concentrations of TSA for defined periods of time. DNA synthesis was assessed by measuring the incorporation of 5-bromo-2'-deoxyuridine. Gene expression at the level of mRNA was quantified by real-time polymerase chain reaction. Expression and phosphorylation of proteins was monitored by immunoblotting, applying an infrared imaging technology. To study the role of p38 MAP kinase, the specific enzyme inhibitor SB202190 and an inactive control substance, SB202474, were employed.
RESULTS: TSA most efficiently inhibited BrdU incorporation in BxPC-3 cells, while CAPAN-1 cells displayed the lowest and AsPC-1 cells an intermediate sensitivity. The biological response of the cell lines correlated with the increase of histone H3 acetylation after TSA application. In BxPC-3 cells (which are wild-type for KRAS), TSA strongly inhibited phosphorylation of ERK 1/2 and AKT. In contrast, activities of ERK and AKT in AsPC-1 and CAPAN-1 cells (both expressing oncogenic KRAS) were not or were only modestly affected by TSA treatment. In all three cell lines, but most pronounced in BxPC-3 cells, TSA exposure induced an activation of the MAP kinase p38. Inhibition of p38 by SB202190 slightly but significantly diminished the antiproliferative effect of TSA in BxPC-3 cells. Interestingly, only BxPC-3 cells responded to TSA treatment by a significant increase of the mRNA levels of bax, a pro-apoptotic member of the BCL gene family. Finally, in BxPC-3 and AsPC-1 cells, but not in the cell line CAPAN-1, significantly higher levels of the cell cycle inhibitor protein p21(Waf1) were observed after TSA application.
CONCLUSION: The biological effect of TSA in PC cells correlates with the increase of acetyl-H3, p21(Waf1), phospho-p38 and bax levels, and the decrease of phospho-ERK 1/2 and phospho-AKT.

• 出版日期2010-4-28