Functional Characterization of a Fluorescent Highly Tumorigenic Ovarian Cancer Line to Test Cellular Therapy in Experimental Models

Abstract

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.

Cellular therapy in combination with cytokines improves survival in a xenograft mouse model of ovarian cancer

“…This cytotoxic response was not observed in the absence of IL-2 indicating that IL-2 is necessary for the PBMC to elicit the cytotoxic response [7,10]. In addition, we have shown that IFNa-2b has an inhibitory effect on ovarian cancer cell growth [7]. Therefore, in this study, we proposed to further evaluate cellular therapy using different combinations of cytokines (IL-2 and IFNa-2b) in combination with PBMC in a xenograft murine model of ovarian cancer in order to study their effects in vivo.…”

“…Cell line culture SKOV3-AF2 cells were derived from SKOV-3 (ATCC, Manassas, VA) cells as recently described by Ingersoll et al [7,8]. Briefly, SKOV3-AF2 cells were derived by first stably transfecting SKOV-3 cells with red-fluorescent protein (SKOV3-RFP), so that the cells could be distinguished from the immune effector cells by fluorescence.…”

“…The line derived from this tumor suspension was called SKOV3-AF2. We have previously shown that these SKOV3-AF2 cells are more tumorigenic in nude mice, show increased resistance to IFNa-2b in vitro, and are sensitive to immune-mediated cytotoxicity in vitro [7,8]. All the cells were grown in McCoy's media (Invitrogen, Carlsbad, CA) supplemented with 10 % fetal bovine serum (FBS; ATCC) and 1 % penicillin and streptomycin (Invitrogen) in 5 % CO 2 at 37°C.…”

“…Briefly, SKOV3-AF2 cells were derived by first stably transfecting SKOV-3 cells with red-fluorescent protein (SKOV3-RFP), so that the cells could be distinguished from the immune effector cells by fluorescence. Next, the cells were passaged through nude mice to make the cells more tumorigenic in a xenograft mouse model as described previously [7,8]. Single-cell suspensions were made from solid tumor and ascitic fluid harvested from the parental SKOV3-RFP tumors and grown in culture.…”

“…Peripheral blood mononuclear cells (PBMC) in the presence of the cytokines [interleukin-2 (IL-2) and interferon a-2b (IFNa-2b)], elicit up to a 70 % cytotoxic response against ovarian cancer cells in vitro [7][8][9][10]. This cytotoxic response was not observed in the absence of IL-2 indicating that IL-2 is necessary for the PBMC to elicit the cytotoxic response [7,10]. In addition, we have shown that IFNa-2b has an inhibitory effect on ovarian cancer cell growth [7].…”