title: Targeting PI3K and RAD51 in Barrett's adenocarcinoma: impact on DNA damage checkpoints, expression profile and tumor growth.
abstract: Phosphatidylinositol 3-kinase (PI3K)/v-akt murine thymoma viral oncogene homolog 1 (AKT) signaling in cancer is implicated in various survival pathways including regulation of recombinase (RAD51). In this study, we evaluated PI3K and RAD51 as targets in Barrett's adenocarcinoma (BAC) cells both in vitro and in vivo. BAC cell lines (OE19, OE33, and FLO-1) were cultured in the presence of PI3K inhibitor (wortmannin) and the impact on growth and expression of AKT, phosphorylated-AKT (P-AKT), and RAD51 was determined. Wortmannin induced growth arrest and apoptosis in two BAC cell lines (OE33 and OE19), which had relatively higher expression of AKT. FLO-1 cells, with lower AKT expression, were less sensitive to treatment and investigated further. In FLO-1 cells, wortmannin suppressed ataxia telangiectasia and Rad3-related protein (ATR)-checkpoint kinase 1 (CHK1)-mediated checkpoint and multiple DNA repair genes, whereas RAD51 and CHK2 were not affected. Western blotting confirmed that RAD51 was suppressed by wortmannin in OE33 and OE19 cells, but not in FLO-1 cells. Suppression of RAD51 in FLO-1 cells down-regulated the expression of CHK2 and CHK1, and reduced the proliferative potential. Finally, the suppression of RAD51 in FLO-1 cells, significantly increased the anticancer activity of wortmannin in these cells, both in vitro and in vivo. We show that PI3K signaling and hsRAD51, through distinct roles in DNA damage response and repair pathways, provide survival advantage to BAC cells. In cells with inherent low expression of AKT, RAD51 is unaffected by PI3K suppression and provides an additional survival pathway. Simultaneous suppression of PI3K and RAD51, especially in cells with lower AKT expression, can significantly reduce their proliferative potential.
authors.affiliation: Department of Adult Oncology, Harvard (Dana Farber) Cancer Institute, Boston, MA, USA.
authors.forename: Jagannath
authors.initials: J
authors.lastname: Pal
authors.forename: Mariateresa
authors.initials: M
authors.lastname: Fulciniti
authors.forename: Puru
authors.initials: P
authors.lastname: Nanjappa
authors.forename: Leutz
authors.initials: L
authors.lastname: Buon
authors.forename: Yu-Tzu
authors.initials: YT
authors.lastname: Tai
authors.forename: Pierfrancesco
authors.initials: P
authors.lastname: Tassone
authors.forename: Nikhil C
authors.initials: NC
authors.lastname: Munshi
authors.forename: Masood A
authors.initials: MA
authors.lastname: Shammas
mesh_terms: D000230:Adenocarcinoma / Q000201:enzymology / Q000235:genetics*; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D016000:Cluster Analysis; D004249:DNA Damage; D004938:Esophageal Neoplasms / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D008297:Male; D051379:Mice; D016513:Mice, SCID; D058539:Phosphatidylinositol 3-Kinase / Q000235:genetics / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051135:Rad51 Recombinase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D000081082:Phosphoinositide-3 Kinase Inhibitors; D034741:RNA, Small Interfering; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; D051135:Rad51 Recombinase; D000077191:Wortmannin