title: Functional characterization of a fluorescent highly tumorigenic ovarian cancer line to test cellular therapy in experimental models.
abstract: OBJECTIVE
  The objective of this study was to functionally characterize a fluorescent highly tumorigenic ovarian cancer line to test cellular therapy in combination with cytokines or chemotherapies in experimental models.
  
  
  METHODS
  A fluorescent highly tumorigenic subline (SKOV3-AF2) was derived from the SKOV3 ovarian cancer cell line. Peripheral blood mononuclear cell (PBMC)-mediated cytotoxicity of SKOV3-AF2 in the presence of interleukin 2 (IL-2) and interferon α-2b (IFNα-2b) was assayed by lactate dehydrogenase release. Sensitivity of SKOV3-AF2 cells to polyethylene glycol-IFNα-2b and IL-2 was assayed in a xenograph nude mouse model. Histopathology was performed to determine necrosis and tumor-infiltrating lymphocytes in the solid tumors. Reverse transcriptase-polymerase chain reaction was used for gene expression analyses of E-cadherin and cysteine-rich 61 (CCN1).
  
  
  RESULTS
  The SKOV3-AF2 subline exhibits increased cytotoxicity (up to 70%), mediated by PBMCs, IL-2, and IFNα-2b, compared with parental SKOV3-red fluorescent protein (RFP) cells. SKOV3-AF2 cells are more tumorigenic in vivo as indicated by tumor incidence, time to sacrifice, tumor weight, and ascitic fluid production. SKOV3-AF2 tumor growth was inhibited by polyethylene glycol-IFNα-2b but not low-dose IL-2. Histopathology revealed that the tumors consisted of poorly differentiated surface epithelial carcinoma. SKOV3-RFP, and -AF2 cell lines as well as -AF2 tumors expressed E-cadherin. SKOV3-AF2 derived tumors expressed CCN1; however, the SKOV3-RFP and SKOV3-AF2 cell lines did not.
  
  
  CONCLUSIONS
  Characterization of SKOV3-AF2 cells revealed that it is more susceptible to PBMC-mediated cytotoxicity than SKOV3-RFP and highly tumorigenic in a xenograph model, and AF-2 tumors express genes that promote aggressive behavior. Collectively, our data suggest that the SKOV3-AF2 subline will be a useful tool to test cellular therapy for the treatment of ovarian cancer utilizing experimental models.
authors: Florida Hospital Gynecologic Oncology, Florida Hospital Cancer Institute, Orlando, FL 32804, USA. susan.blaydes@flhosp.org
authors: Susan Blaydes
authors: SB
authors: Ingersoll
authors: Sarfraz
authors: S
authors: Ahmad
authors: Gregory P
authors: GP
authors: Stoltzfus
authors: Sheylan
authors: S
authors: Patel
authors: Michael J
authors: MJ
authors: Radi
authors: Neil J
authors: NJ
authors: Finkler
authors: John R
authors: JR
authors: Edwards
authors: Robert W
authors: RW
authors: Holloway
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D005260:Female; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006801:Humans; D007376:Interleukin-2 / Q000627:therapeutic use*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism; D016246:Lymphocytes, Tumor-Infiltrating / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D010051:Ovarian Neoplasms / Q000473:pathology* / Q000517:prevention & control*
chemical_list: D007376:Interleukin-2; D049452:Green Fluorescent Proteins