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Published on: September 30, 2016
Antitumor activity of iNGR-GRIM-19 in colorectal cancer
Li Pang1, Yan Xia2, Dawei Wang1
1Department of Emergency, The First Hospital of Jilin University.
Background:
Gene associated with retinoid-interferon induced mortality-19 (GRIM-19) plays crucial roles in carcinogenesis.
Objective:
To explore the antitumor activity of internalizing NGR (iNGR) gene associated with GRIM-19 in colorectal cancer.
Methods:
Cells were incubated with fluorescein isothiocyanate-labeled fusion proteins followed by fluorescence microscopic analysis. Cell proliferation was determined by MTT assay. Cell cycle was analyzed by flow cytometric analysis. Cell migration and invasion capacity were evaluated by wound scratch and Transwell assays, respectively. Apoptosis was measured by Annexin V/PI staining and TUNEL assay. Gene expressions were determined by RT-PCR and Western blotting. Nude mice bearing colorectal cancer received vehicle, GRIM-19, or iNGR-GRIM-19 fusion protein injection, and the in vivo antitumor capacity of the fusion proteins was examined.
Results:
iNGR-GRIM-19 was specifically taken up by human colorectal cancer Colo205 cells, but not corneal epithelial (HCEpic) cells, whereas GRIM-19 was not internalized by either cell type. Unlike GRIM-19, incubation with iNGR-GRIM-19 dose-dependently inhibited proliferation, induced G1 phase arrest, suppressed cell migration and invasion, and caused apoptosis in Colo205 cells. Additionally, injection of iNGR-GRIM-19 extended the lifespan of colorectal cancer-bearing nude mice and reduced in vivo tumor growth as compared with vehicle or GRIM-19 treatment. iNGR-GRIM-19 was localized only in the tumor mass, without affecting other tissues, such as liver or kidney. iNGR-GRIM-19 injection led to G1 phase arrest and induced cell apoptosis in xenografted colorectal cancer tissues.
Conclusions:
iNGR-GRIM-19 has an efficient antitumor activity in vitro and in vivo, and might be a promising agent for the treatment of colorectal cancer.
Insights
The novel internalizing NGR (iNGR)-GRIM-19 fusion protein demonstrates significant antitumor effects against colorectal cancer cells in vitro and in vivo. This targeted therapy effectively inhibits tumor growth and enhances survival, showing promise for colorectal cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- Gene associated with retinoid-interferon induced mortality-19 (GRIM-19) is implicated in cancer development.
- Targeted delivery of therapeutic agents is crucial for effective cancer treatment.
Purpose of the Study:
- To evaluate the antitumor activity of the internalizing NGR (iNGR)-GRIM-19 fusion protein in colorectal cancer.
- To assess the specificity and efficacy of iNGR-GRIM-19 in preclinical models.
Main Methods:
- Utilized cell culture assays (MTT, flow cytometry, wound scratch, Transwell, Annexin V/PI, TUNEL) to assess proliferation, cell cycle, migration, invasion, and apoptosis.
- Employed RT-PCR and Western blotting for gene expression analysis.
- Investigated in vivo antitumor efficacy in nude mice bearing colorectal cancer xenografts.
Main Results:
- iNGR-GRIM-19 selectively targeted and was internalized by colorectal cancer cells, unlike GRIM-19 alone.
- iNGR-GRIM-19 significantly inhibited proliferation, induced G1 phase arrest, suppressed migration and invasion, and promoted apoptosis in vitro.
- In vivo studies showed iNGR-GRIM-19 reduced tumor growth, extended survival, and exhibited tumor-specific localization without affecting healthy organs.
Conclusions:
- iNGR-GRIM-19 exhibits potent in vitro and in vivo antitumor activity against colorectal cancer.
- The targeted delivery via iNGR enhances the therapeutic potential of GRIM-19.
- iNGR-GRIM-19 represents a promising candidate for future colorectal cancer therapies.
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