title: In vitro studies on the modification of low-dose hyper-radiosensitivity in prostate cancer cells by incubation with genistein and estradiol.
abstract: BACKGROUND
  As the majority of prostate cancers (PC) express estrogen receptors, we evaluated the combination of radiation and estrogenic stimulation (estrogen and genistein) on the radiosensitivity of PC cells in vitro.
  
  
  METHODS
  PC cells LNCaP (androgen-sensitive) and PC-3 (androgen-independent) were evaluated. Estrogen receptor (ER) expression was analyzed by means of immunostaining. Cells were incubated in FCS-free media with genistein 10 microM and estradiol 10 microM 24 h before irradiation and up to 24 h after irradiation. Clonogenic survival, cell cycle changes, and expression of p21 were assessed.
  
  
  RESULTS
  LNCaP expressed both ER-alpha and ER-beta, PC-3 did not. Incubation of LNCaP and PC-3 with genistein resulted in a significant reduction of clonogenic survival. Incubation with estradiol exhibited in low concentrations (0.01 microM) stimulatory effects, while higher concentrations did not influence survival. Both genistein 10 microM and estradiol 10 microM increased low-dose hyper-radiosensitivity [HRS] in LNCaP, while hormonal incubation abolished HRS in PC-3. In LNCaP cells hormonal stimulation inhibited p21 induction after irradiation with 4 Gy. In PC-3 cells, the proportion of cells in G2/M was increased after irradiation with 4 Gy.
  
  
  CONCLUSIONS
  We found an increased HRS to low irradiation doses after incubation with estradiol or genistein in ER-alpha and ER-beta positive LNCaP cells. This is of high clinical interest, as this tumor model reflects a locally advanced, androgen dependent PC. In contrast, in ER-alpha and ER-beta negative PC-3 cells we observed an abolishing of the HRS to low irradiation doses by hormonal stimulation. The effects of both tested compounds on survival were ER and p53 independent. Since genistein and estradiol effects in both cell lines were comparable, neither ER- nor p53-expression seemed to play a role in the linked signalling. Nevertheless both compounds targeted the same molecular switch. To identify the underlying molecular mechanisms, further studies are needed.
authors.affiliation: Department of Radiotherapy and Radiooncology, University hospital Goettingen, Robert-Koch-Str, 40, 37075, Goettingen, Germany. ro.hermann@t-online.de
authors.forename: Robert Michael
authors.initials: RM
authors.lastname: Hermann
authors.forename: Hendrik Andreas
authors.initials: HA
authors.lastname: Wolff
authors.forename: Hubertus
authors.initials: H
authors.lastname: Jarry
authors.forename: Paul
authors.initials: P
authors.lastname: Thelen
authors.forename: Carsten
authors.initials: C
authors.lastname: Gruendker
authors.forename: Margret
authors.initials: M
authors.lastname: Rave-Fraenk
authors.forename: Heinz
authors.initials: H
authors.lastname: Schmidberger
authors.forename: Hans
authors.initials: H
authors.lastname: Christiansen
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003131:Combined Modality Therapy; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000187:drug effects / Q000378:metabolism; D004958:Estradiol / Q000494:pharmacology*; D019833:Genistein / Q000494:pharmacology*; D006801:Humans; D008297:Male; D009376:Neoplasms, Hormone-Dependent / Q000188:drug therapy / Q000532:radiotherapy; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000532:radiotherapy*; D011836:Radiation Tolerance / Q000187:drug effects*; D011960:Receptors, Estrogen / Q000187:drug effects; D014410:Tumor Stem Cell Assay
chemical_list: D000970:Antineoplastic Agents; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D011960:Receptors, Estrogen; D004958:Estradiol; D019833:Genistein