title: Mitochondrial DNA depletion induces radioresistance by suppressing G2 checkpoint activation in human pancreatic cancer cells.
abstract: We hypothesized that mitochondrial function regulates cell cycle checkpoint activation and radiosensitivity. Human pancreatic tumor cells (MiaPaCa-2, rho(+)) were depleted of mitochondrial DNA (rho degrees ) by culturing cells in the presence of ethidium bromide. Depletion of mitochondrial DNA was verified by PCR amplification of total DNA using primer pairs specific for mitochondrial DNA. Loss of mitochondrial DNA decreased plating efficiency and the percentage of cells in S phase. Exponential cultures were irradiated with 2, 4 and 6 Gy (dose rate: 0.83 Gy/min) of ionizing radiation and harvested for determination of cell viability, growth and cell cycle phase distributions. Rho degrees cells were radioresistant compared to rho(+) cells, with a dose-modifying factor (DMF) of 1.6. Although cell growth was significantly inhibited in irradiated rho(+) cells compared to unirradiated control cells, the inhibition in Rho degrees cells was minimal. In addition, mitochondrial DNA depletion suppressed radiation-induced G(2) checkpoint activation, which was accompanied by increases in both cyclin B1 and CDK1. These results suggest that mitochondrial function may regulate cell cycle checkpoint activation and radiosensitivity in pancreatic cancer cells.
authors: Departments of Surgery, Iowa City, Iowa, USA.
authors: Carla R
authors: CR
authors: Cloos
authors: David H
authors: DH
authors: Daniels
authors: Amanda
authors: A
authors: Kalen
authors: Katee
authors: K
authors: Matthews
authors: Juan
authors: J
authors: Du
authors: Prabhat C
authors: PC
authors: Goswami
authors: Joseph J
authors: JJ
authors: Cullen
mesh_terms: D016203:CDC2 Protein Kinase / Q000032:analysis; D045744:Cell Line, Tumor; D019926:Cyclin B / Q000032:analysis; D056744:Cyclin B1; D004272:DNA, Mitochondrial / Q000502:physiology*; D016195:G2 Phase / Q000528:radiation effects*; D006801:Humans; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology / Q000532:radiotherapy*; D011836:Radiation Tolerance
chemical_list: C534699:CCNB1 protein, human; D019926:Cyclin B; D056744:Cyclin B1; D004272:DNA, Mitochondrial; D016203:CDC2 Protein Kinase