title: Characterization of a membrane-active anti-tumor agent, UA8967.
abstract: Deletions or mutations in the tumor suppressor gene DPC4 (deleted in pancreatic carcinoma locus 4) are common in colon and pancreatic cancers. Using the Target-related Affinity Profiling (TRAP) chemical library screening method, a novel agent, UA8967, was selected for further studies because it showed greater potency in DPC4-deleted HCT-116 colon cancer cells. Cytotoxicity studies in six pancreatic cancer cell lines (MiaPaca-2, Panc-1, BxPC3, CF-PAC1, AsPC1, and T3M4), one normal human pancreatic ductal epithelial line (HPDE-6) and the HCT-116 DPC4(+/+) and HCT-116 DPC4(-/-) colon cancer cells showed IC50s ranging from 12-61 μM for exposure times of 72 h. Analysis of schedule dependence showed no advantage for long drug exposure times. There was also no selective inhibition of DNA, RNA or protein synthesis after exposure to UA8967. At 24-48 h, there was an accumulation of cells in G0/G1-phase and a proportionate reduction in S-phase cells. Within 1-6 h of exposure, cells were found to undergo an autophagic response, followed at 24 h by a low level of caspase-independent apoptosis with some necrosis. Because of the relatively non-specific mechanistic effects of UA8967, plasma membrane viability was evaluated using uptake of trypan blue and Sytox® Green dyes, and leakage of LDH. There was a dose dependent increase in Sytox® Green staining, trypan blue uptake and LDH leakage with increasing concentrations of UA8967, suggesting that UA8967 is affecting the plasma membrane. The DPC4(-/-) cells were more sensitive to UA8967 but not to DMSO, suggesting a drug-specific effect on cell membrane integrity.
authors.affiliation: University of Arizona Cancer Center, 1515 N. Campbell Ave, Rm 4963C, Tucson, AZ 85724-5024, USA. bdorr@azcc.arizona.edu
authors.forename: Robert T
authors.initials: RT
authors.lastname: Dorr
authors.forename: Betty K
authors.initials: BK
authors.lastname: Samulitis
authors.forename: Lee
authors.initials: L
authors.lastname: Wisner
authors.forename: Haiyong
authors.initials: H
authors.lastname: Han
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhao
authors.forename: Paul
authors.initials: P
authors.lastname: Beroza
authors.forename: Komath
authors.initials: K
authors.lastname: Damodaran
authors.forename: Suzu
authors.initials: S
authors.lastname: Igarashi
authors.forename: Terry H
authors.initials: TH
authors.lastname: Landowski
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Von Hoff
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D007211:Indoles / Q000494:pharmacology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010879:Piperazines / Q000494:pharmacology*; D051901:Smad4 Protein / Q000235:genetics*
chemical_list: C584560:3-((4-benzylpiperazino)methyl)-1H-indole; D000970:Antineoplastic Agents; D007211:Indoles; D010879:Piperazines; C497796:SMAD4 protein, human; D051901:Smad4 Protein; D007770:L-Lactate Dehydrogenase