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

2011 ◽  
Vol 21 (3) ◽  
pp. 457-465 ◽  
Author(s):  
Susan Blaydes Ingersoll ◽  
Sarfraz Ahmad ◽  
Gregory P. Stoltzfus ◽  
Sheylan Patel ◽  
Michael J. Radi ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Polyethylene Glycol ◽  
Cell Lines ◽  
Cellular Therapy ◽  
Functional Characterization ◽  
Interleukin 2 ◽  
Experimental Models ◽  
Cancer Line ◽  
E Cadherin

ObjectivesThe 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.MethodsA 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 ofE-cadherinandcysteine-rich 61(CCN1).ResultsThe 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 expressedE-cadherin.SKOV3-AF2 derived tumors expressedCCN1; however, the SKOV3-RFP and SKOV3-AF2 cell lines did not.ConclusionsCharacterization 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.

Abstract 2708: Functional characterization of a fluorescent highly tumorigenic ovarian cancer line to test cellular therapy in experimental models

2011 ◽  
Author(s):  
Susan Blaydes Ingersoll ◽  
Sarfraz Ahmad ◽  
Gregory P. Stoltzfus ◽  
Mohammed H. Merchant ◽  
Michael J. Radi ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cellular Therapy ◽  
Functional Characterization ◽  
Experimental Models ◽  
Cancer Line

scholarly journals NFB-01. FUNCTIONAL CHARACTERIZATION OF ATRX LOSS IN NF1-ASSOCIATED GLIOMA AND MPNST

2020 ◽  
Vol 22 (Supplement_3) ◽  
pp. iii417-iii418
Author(s):  
Ming Yuan ◽  
Karlyne Reilly ◽  
Christine Pratilas ◽  
Christopher Heaphy ◽  
Fausto Rodriguez
Keyword(s):  
Cell Lines ◽  
Functional Characterization ◽  
Tert Promoter ◽  
Aggressive Tumors ◽  
Sirna Knockdown ◽  
Nih 3T3 ◽  
Vitro Effect ◽  
Murine Glioma

Abstract To identify the biologic relevance of ATRX loss in NF1-associated gliomagenesis, we studied the effects of Atrx loss using four previously characterized Nf1+/-Trp53+/- murine glioma lines. Lines 130G#3 and 158D#8 (corresponding to grade IV and III gliomas, respectively) displayed preserved ATRX protein expression compared to NIH-3T3 cells. We studied the effects of Atrx knockdown in these two lines in the presence and absence of the TERT inhibitor, BIRBR1532. Using a telomere-specific FISH assay, we identified increased signal intensity after Atrx knockdown, only in the presence of the TERT inhibitor. These features are reminiscent of ALT, although there were no significant alterations in cell growth. Next, we studied the effect of ATRX loss in MPNST lines ST88-14, NF90-8, STS-26T. These cell lines all expressed ATRX and DAXX. However, STS-26T contained a TERT promoter mutation and ST88-14 had a known SNP in the TERT promoter, while NF90-8 had no alterations. ATRX siRNA knockdown showed no significant effects in cell proliferation or apoptosis. However, ATRX knockdown resulted in rare ultra-bright foci, indicative of ALT. Next, we studied the in vitro effect of the ATR inhibitor VE-821 in MPNST cell lines. Only NF90-8 (lacking TERT alterations) demonstrated a decrease in growth after ATRX knockdown and VE-821 treatment. However, ATRX knockdown alone did not affect sensitivity to carboplatin. Our findings further support a role for ATRX loss with subsequent ALT activation in a biologic subset of NF1-associated malignancies, thereby opening an opportunity for therapeutic targeting of these aggressive tumors using specific classes of drugs.

In vitro phenotypic and functional characterization of human pigment epithelial cell lines

1989 ◽  
Vol 8 (5) ◽  
pp. 435-440 ◽  
Author(s):  
Kamla Dutt ◽  
J. Clifford Waldrep ◽  
Henry J. Kaplan ◽  
Monte Del Monte ◽  
Eugene Semple ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Lines ◽  
Functional Characterization ◽  
Pigment Epithelial ◽  
Epithelial Cell Lines

scholarly journals Biochemical and biological characterization of lymphocyte regulatory molecules. V. Identification of an interleukin 2-producing human leukemia T cell line.

1980 ◽  
Vol 152 (6) ◽  
pp. 1709-1719 ◽  
Author(s):  
S Gillis ◽  
J Watson
Keyword(s):  
T Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Human Peripheral Blood ◽  
Fatty Acid Derivative ◽  
Interleukin 2 ◽  
Leukemia Cell ◽  
Tumor Cell Line ◽  
Human Leukemia

To isolate a stable tumor cell line capable of producing human interleukin 2 (IL-2; formerly referred to as T cell growth factor), 16 human T and B leukemia cell lines were screened for constitutive and mitogen-stimulated IL-2 production. We found that the T cell leukemia line designated Jurkat-FHCRC produced > 200 U/ml of IL-2 activity after a 24-h stimulation with T cell mitogens. Peak mitogen-induced IL-2 activity was found in supernates harvested from 24-h Jurkat-FHCRC cell cultures stimulated with either 1% phytohemagglutinin or 20 microgram/ml concanavalin A. Addition of the fatty acid derivative phorbol myristate acetate to mitogen-stimulated cultures increased Jurkat-FHCRC IL-2 production to concentrations > 400 U/ml. IL-2 activity observed in such cases represented between 100--300 times that produced in conventional cultures of mitogen- or alloantigen-stimulated normal human peripheral blood or splenic lymphocytes. Jurkat-FHCRC-derived conditioned medium demonstrated equal capacity to promote the sustained in vitro proliferation of either murine or human activated T cell lines confirming the ability of Jurkat-FHCRC cells to produce human IL-2. These studies identify a new source of human IL-2 and establish a valuable reagent for the isolation and further molecular characterization of this immunoregulatory molecule.

Sign in / Sign up

Export Citation Format

Share Document