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Evaluation of Stem Cell Properties in Human Ovarian Carcinoma Cells Using Multi and Single Cell-based Spheres Assays
Published on: January 3, 2015
EGF-related antisense oligonucleotides inhibit the proliferation of human ovarian carcinoma cells
A Casamassimi1, A De Luca, S Agrawal
1Novel Therapeutic Approaches Section-Oncologia Sperimentale D, ITN-Fondazione Pascale, Napoli, Italy.
Background:
The epidermal growth factor (EGF)-like peptides CRIPTO (CR), amphiregulin (AR) and transforming growth factor alpha (TGFalpha) are expressed in human ovarian carcinomas.
Materials And Methods:
The expression of AR, CR and TGFalpha in ovarian carcinoma cell lines was assessed by immunocytochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). The antiproliferative effects of antisense phosphorothioate oligodeoxynucleotides (AS S-Oligos) directed against either AR, CR or TGFalpha was evaluated by using a clonogenic assay.
Results:
A majority of the ovarian carcinoma cell lines was found to express TGFalpha, AR and CR mRNAs and proteins. AS S-Oligos directed against either AR, CR or TGFalpha were able to inhibit the anchorage-independent growth of NIH:OVCAR3 and NIH:OVCAR8 cells in a dose dependent manner. A 30%-50% growth inhibition was observed at a 2 microM concentration of the AS S-Oligos. Treatment of these cells with combinations of EGF-related AS S-Oligos resulted in a more significant growth inhibition when compared to treatment with a single AS S-oligo. A 60%-75% growth inhibition was observed using combinations of AR, CR and TGFalpha AS S-oligos at a total concentration of 2 microM. An additive growth-inhibitory effect occurred when ovarian carcinoma cells were exposed to the AS S-Oligos after treatment with either paclitaxel or cis-platinum.
Conclusions:
These data suggest that EGF-related peptides function as autocrine growth factors in ovarian carcinoma cells, and that they might represent targets for experimental therapy of ovarian carcinoma.
Insights
Antisense oligodeoxynucleotides targeting epidermal growth factor-like peptides CRIPTO, amphiregulin, and transforming growth factor alpha inhibit ovarian carcinoma cell growth. Combining these treatments with chemotherapy enhances their antiproliferative effects, suggesting novel therapeutic strategies.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Epidermal growth factor (EGF)-like peptides, including CRIPTO (CR), amphiregulin (AR), and transforming growth factor alpha (TGFalpha), are expressed in human ovarian carcinomas.
- These peptides are implicated in cancer cell proliferation and survival.
Purpose of the Study:
- To investigate the expression of AR, CR, and TGFalpha in ovarian carcinoma cell lines.
- To evaluate the antiproliferative effects of antisense oligodeoxynucleotides (AS S-Oligos) targeting these EGF-related peptides.
- To explore the potential of these peptides as therapeutic targets in ovarian cancer.
Main Methods:
- Immunocytochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) were used to assess peptide expression.
- Clonogenic assays evaluated the effects of AS S-Oligos on anchorage-independent growth.
- Combinatorial treatments with AS S-Oligos and chemotherapy agents (paclitaxel, cis-platinum) were analyzed.
Main Results:
- Ovarian carcinoma cell lines predominantly expressed AR, CR, and TGFalpha at both mRNA and protein levels.
- AS S-Oligos targeting AR, CR, or TGFalpha significantly inhibited cancer cell growth in a dose-dependent manner.
- Combined AS S-Oligos demonstrated enhanced growth inhibition, and additive effects were observed when used with paclitaxel or cis-platinum.
Conclusions:
- EGF-related peptides function as autocrine growth factors in ovarian carcinoma.
- Targeting these peptides with AS S-Oligos shows promise for experimental ovarian cancer therapy.
- Combination therapies involving these agents and chemotherapy may offer improved treatment outcomes.
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