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Inhibition of CRIPTO expression and tumorigenicity in human colon cancer cells by antisense RNA and
F Ciardiello1, G Tortora, C Bianco
1Cattedra di Oncologia Medica, Facoltà di Medicina e Chirurgia, Università degli Studi di Napoli Federico II, Italy.
Abstract:
CRIPTO is an epidermal growth factor-related gene expressed in a majority of human colorectal tumors. To assess the role of CRIPTO in the growth control of human colon cancer, we have treated human colon carcinoma GEO and CBS cells, that possess high levels of CRIPTO, and WIDR colon cancer cells, that are negative for CRIPTO expression, with two antisense phosphorothioate oligodeoxynucleotides complementary to the 5' end of the human CRIPTO mRNA. Both antisense oligodeoxynucleotides significantly reduced endogenous CRIPTO protein levels and inhibited GEO and CBS cell growth in monolayer and in semisolid medium, whereas they did not affect WIDR cell growth. In addition, GEO, CBS and WIDR cells were infected with a recombinant retroviral vector containing the hygromycin-resistance gene and a 900 bp EcoRI-EcoRI coding fragment of the human CRIPTO cDNA oriented in the 3' to 5' direction. GEO and CBS CRIPTO antisense infectants exhibited a 60 to 70% reduction in CRIPTO protein expression, in monolayer growth and in soft agar cloning efficiency as compared to parental noninfected cells. In contrast, infection of WIDR cells with the CRIPTO antisense retrovirus did not alter their growth. Finally, GEO CRIPTO antisense infectants formed tumors in nude mice that were significantly smaller and had a larger latency period as compared to noninfected GEO cells.
Insights
CRIPTO, a gene in colorectal tumors, was targeted using antisense oligodeoxynucleotides. Inhibiting CRIPTO reduced tumor cell growth and tumor size in mice, suggesting CRIPTO
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- CRIPTO is an epidermal growth factor-related gene.
- It is expressed in most human colorectal tumors.
- CRIPTO's role in colon cancer growth control requires investigation.
Purpose of the Study:
- To investigate the role of CRIPTO in human colon cancer growth.
- To assess the therapeutic potential of targeting CRIPTO.
Main Methods:
- Utilized antisense phosphorothioate oligodeoxynucleotides targeting human CRIPTO mRNA.
- Infected colon cancer cells (GEO, CBS, WIDR) with CRIPTO antisense retroviral vectors.
- Assessed cell growth in monolayer, semisolid medium, and soft agar.
- Evaluated tumor formation in nude mice.
Main Results:
- Antisense treatment significantly reduced CRIPTO protein levels in GEO and CBS cells.
- Inhibition of CRIPTO reduced growth of GEO and CBS cells but not WIDR cells.
- CRIPTO antisense infectants showed reduced growth and cloning efficiency.
- Tumors formed by CRIPTO antisense GEO cells were smaller with longer latency in nude mice.
Conclusions:
- CRIPTO plays a significant role in colon cancer cell growth.
- Targeting CRIPTO with antisense strategies can inhibit tumor progression.
- CRIPTO is a potential therapeutic target for colorectal cancer.