title: In vitro anticancer effects of a novel immunostimulant: keyhole limpet hemocyanin.
abstract: BACKGROUND
  Keyhole limpet hemocyanin (KLH) is a recently described immune stimulant and hapten carrier derived from a circulating glycoprotein of the marine mollusk Megathura crenulata. It has been reported to be a potent form of intravesical immunotherapy for the treatment of transitional cell carcinoma of the bladder and has been used in a variety of genitourinary tumors. We hypothesized that KLH would be effective against other cancer cells in vitro.
  
  
  METHODS
  Multiple cancer cell lines were tested, including estrogen-dependent breast (MCF-7), estrogen-independent breast (ZR75-1), pancreas (PANC-1, MIA-PaCa), and prostate (DU145). Serial twofold dilutions of KLH were prepared in sterile 96-well plates. Dose-response curves were performed beginning with a concentration of 100 microg of KLH/well and ending at a concentration of 0.8 ng/well. Cells were added at concentrations of 5 x 10(4) cells per well. Cell viability was evaluated at 24 and 72 h by MTT assay at an absorbance of 570 nm.
  
  
  RESULTS
  Significant (P < 0.05) cancer cell growth inhibition was observed in four of the five cell lines tested at both time treatment intervals. The breast cancer line ZR75-1 exhibited a mean growth inhibition of 43 +/- 1.1% (range 37 to 59%) at 72 h, whereas treated MCF-7 cells had an average of 39 +/- 9.1% growth inhibition (range 35 to 44%) at these same concentrations. Treated PANC-1 cells had a mean growth inhibition of 19 +/- 0.8% (range 4 to 46%) at 72 h. The DU145 prostate cancer cell line averaged a 6 +/- 1.3% growth inhibition (range -19 to 55%) over the concentrations tested.
  
  
  CONCLUSIONS
  The direct growth inhibition of multiple tumor cell-lines exhibited by KLH is significant and warrants further in vitro mechanistic studies and in vivo experiments. Investigation into the efficacy and mechanism of response could directly lead to more effective treatment regimens for patients suffering from these diseases.
authors.affiliation: Department of Surgery, West Virginia University, PO Box 9238, Morgantown, 26506, USA.
authors.forename: Dale R
authors.initials: DR
authors.lastname: Riggs
authors.forename: Barbara
authors.initials: B
authors.lastname: Jackson
authors.forename: Linda
authors.initials: L
authors.lastname: Vona-Davis
authors.forename: David
authors.initials: D
authors.lastname: McFadden
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000627:therapeutic use*; D000368:Aged; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000473:pathology; D002455:Cell Division / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005260:Female; D006433:Hemocyanins / Q000008:administration & dosage / Q000627:therapeutic use*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D000276:Adjuvants, Immunologic; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin