Related Experiment Video
Updated: Feb 25, 2026
In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice
Published on: August 23, 2019
Silencing Intersectin 1 Slows Orthotopic Neuroblastoma Growth in Mice
Jamie Harris1, Erika Herrero-Garcia, Angela Russo
1*Department of Surgery, Rush University Medical Center †Department of Pharmacology, College of Medicine, University of Illinois at Chicago ‡Jesse Brown VA Medical Center Departments of § Pathology ¶Surgery, University of Illinois at Chicago ∥University of Illinois Cancer Center, University of Illinois at Chicago, Chicago, IL.
Abstract:
Neuroblastoma accounts for 15% of all pediatric cancer deaths. Intersectin 1 (ITSN1), a scaffold protein involved in phosphoinositide 3-kinase (PI3K) signaling, regulates neuroblastoma cells independent of MYCN status. We hypothesize that by silencing ITSN1 in neuroblastoma cells, tumor growth will be decreased in an orthotopic mouse tumor model. SK-N-AS neuroblastoma cells transfected with empty vector (pSR), vectors expressing scrambled shRNA (pSCR), or shRNAs targeting ITSN1 (sh#1 and sh#2) were used to create orthotopic neuroblastoma tumors in mice. Volume was monitored weekly with ultrasound. End-point was tumor volume >1000 mm. Tumor cell lysates were analyzed with anti-ITSN1 antibody by Western blot. Orthotopic tumors were created in all cell lines. Twenty-five days post injection, pSR tumor size was 917.6±247.7 mm, pSCR was 1180±159.9 mm, sh#1 was 526.3±212.8 mm, and sh#2 was 589.2±74.91 mm. sh#1-tumors and sh#2-tumors were smaller than pSCR (P=0.02), no difference between sh#1 and sh#2. Survival was superior in sh#2-tumors (P=0.02), trended towards improved survival in sh#1-tumors (P=0.09), compared with pSCR-tumors, no difference in pSR tumors. Western blot showed decreased ITSN1 expression in sh#1 and sh#2 compared with pSR and pSCR. Silencing ITSN1 in neuroblastoma cells led to decreased tumor growth in an orthotopic mouse model. Orthotopic animal models can provide insight into the role of ITSN1 pathways in neuroblastoma tumorigenesis.
Insights
Silencing Intersectin 1 (ITSN1) in neuroblastoma cells significantly reduced tumor growth in a mouse model. This finding highlights ITSN1 as a potential therapeutic target for pediatric neuroblastoma.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Neuroblastoma is a significant cause of cancer mortality in children.
- Intersectin 1 (ITSN1), a scaffold protein, influences phosphoinositide 3-kinase (PI3K) signaling in neuroblastoma cells.
- ITSN1's role in neuroblastoma progression is independent of MYCN amplification status.
Purpose of the Study:
- To investigate the effect of silencing ITSN1 on neuroblastoma tumor growth.
- To evaluate ITSN1 as a potential therapeutic target in neuroblastoma.
Main Methods:
- SK-N-AS neuroblastoma cells were transfected with vectors targeting ITSN1 (sh#1, sh#2) or control vectors (pSR, pSCR).
- Orthotopic tumors were established in mice using these cell lines.
- Tumor volume was monitored via ultrasound, and ITSN1 expression was analyzed by Western blot.
Main Results:
- Silencing ITSN1 (sh#1, sh#2) resulted in significantly smaller tumors compared to controls (pSCR).
- Tumors with silenced ITSN1 showed decreased growth rates.
- Survival analysis indicated improved outcomes in mice with ITSN1-silenced tumors.
Conclusions:
- Silencing ITSN1 effectively reduces neuroblastoma tumor growth in an orthotopic mouse model.
- ITSN1 is a promising therapeutic target for neuroblastoma.
- Orthotopic models are valuable for studying ITSN1's role in neuroblastoma tumorigenesis.
