Abstract | PURPOSE: EXPERIMENTAL DESIGN:
Tumor cells obtained from primary cultures of five ovarian and one cervical tumor were mixed with an equal proportion of NIH-3T3 fibroblasts and plated inside collagen gels in tissue culture plates. After 4-7 days of growth, these organotypic cultures were treated with media alone, SS1(dsFv)PE38, and a control immunotoxin RFB4(dsFv)PE38, which targets the CD22 antigen not present on gynecologic tumors, every other day x 3. The organotypic culture gels were then formalin fixed, paraffin embedded, and evaluated for immunotoxin sensitivity using light microscopic examination of H&E-stained slides and also evaluated for apoptosis using the terminal deoxynucleotidyl transferase-mediated nick end labeling assay. RESULTS:
Tumors expressing mesothelin showed a significant dose-dependent sensitivity to SS1(dsFv)PE38 even at concentrations as low as 1 ng/ml, whereas no antitumor activity was seen at 100 ng/ml in tumors that did not express mesothelin. This activity was specifically attributable to mesothelin targeting because RFB4 (dsFv)-PE38 had no activity against mesothelin-expressing tumors. CONCLUSIONS: These results demonstrate that ovarian and cervical tumor cells obtained from patients can be grown in short-term culture using an organotypic culture model. Our results also show low concentrations of an immunotoxin targeting mesothelin is cytotoxic to mesothelin-expressing human tumors by inducing apoptosis.
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Authors | Raffit Hassan, Megan R Lerner, Doris Benbrook, Stan A Lightfoot, Daniel J Brackett, Qing-Cheng Wang, Ira Pastan |
Journal | Clinical cancer research : an official journal of the American Association for Cancer Research
(Clin Cancer Res)
Vol. 8
Issue 11
Pg. 3520-6
(Nov 2002)
ISSN: 1078-0432 [Print] United States |
PMID | 12429643
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antibodies, Monoclonal
- Enterotoxins
- GPI-Linked Proteins
- Immunoglobulin Fragments
- Immunotoxins
- Membrane Glycoproteins
- Msln protein, mouse
- Proteins
- Recombinant Proteins
- SS(dsFv)PE38
- SS1(dsFv)PE38
- immunoglobulin Fv
- Collagen
- Mesothelin
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Topics |
- 3T3 Cells
- Animals
- Antibodies, Monoclonal
- Apoptosis
- Cells, Cultured
- Coculture Techniques
- Collagen
(pharmacology)
- Dose-Response Relationship, Drug
- Enterotoxins
- Female
- Fibroblasts
(metabolism)
- GPI-Linked Proteins
- Genital Neoplasms, Female
(drug therapy)
- Humans
- Immunoglobulin Fragments
(genetics, pharmacology)
- Immunohistochemistry
- Immunotoxins
(pharmacology)
- In Situ Nick-End Labeling
- Membrane Glycoproteins
(antagonists & inhibitors)
- Mesothelin
- Mice
- Proteins
- Recombinant Proteins
- Tumor Cells, Cultured
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