Silencing of the MYCN gene by siRNA delivered by folate receptor-targeted liposomes in LA-N-5 cells
- Chen FengAffiliated withDepartment of Pediatrics, Chinese PLA General Hospital
- , Tianyou WangAffiliated withDepartment of Hematology-Oncology, Capital Institute of Pediatrics
- , Ruihong TangAffiliated withDepartment of Hematology-Oncology, Capital Institute of Pediatrics
- , Jianwen WangAffiliated withDepartment of Pediatrics, Chinese PLA General Hospital
- , Hui LongAffiliated withDepartment of Pediatrics, Chinese PLA General Hospital
- , Xiaoning GaoAffiliated withDepartment of Pediatrics, Chinese PLA General Hospital
- , Suoqin TangAffiliated withDepartment of Pediatrics, Chinese PLA General Hospital Email author
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MYCN amplification is highly associated with malignancy and correlates with poor prognosis in patients with neuroblastoma.
Materials and methods
We developed a novel liposome-MYCN siRNA-folic acid complex, and the transfection efficacy was measured in LA-N-5 cells by cy-3 fluorescence density in each microgram of protein from the transfected cell lysate. MYCN expression and cell growth were studied with quantitative RT-PCR and MTT assays, and the expression of MYCN protein was studied with Western blot, respectively. An SCID mouse model with subcutaneous LA-N-5 xenografted tumor was established. The animals were divided into four groups (n = 5) and they were peritoneally injected with liposome-encapsulated MYCN siRNA (siRNA 125 μg/kg/day), lipid-encapsulated control siRNA, MYCN siRNA, or liposome only, respectively, for 5 consecutive days. The animals were killed 24 h after the last injection, and the expression of MYCN mRNA in tumor tissue was detected by RT-PCR.
Our results are as follows: the transfect efficacy reached 1808.5 ± 140.2 pg siRNA/μg protein in LA-N-5 lysates after treatment with 100 nmol/L MYCN siRNA encapsulated with lipid, and fluorescence could be visualized in 92% of LA-N-5 cells after transfection. At 72 h post-transfection, MYCN mRNA expression in LA-N-5 cells was downregulated by 79.2%, MYCN protein was downregulated by 71.3% and cell growth was inhibited by 66.2%, as measured by MTT assay. In the in vivo study, MYCN mRNA expression was knocked down 53.1% in tumor tissues with injection of liposome-encapsulated MYCN siRNA as compared to control siRNA.
These results suggest that targeted delivery of MYCN siRNA by folate receptor-targeted lipid vesicles into LA-N-5 cells is efficacious and capable of suppressing MYCN mRNA expression both in vitro and in vivo.
KeywordsNeuroblastoma Target therapy MYCN gene Folate receptor Liposome
- Silencing of the MYCN gene by siRNA delivered by folate receptor-targeted liposomes in LA-N-5 cells
Pediatric Surgery International
Volume 26, Issue 12 , pp 1185-1191
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- Target therapy
- MYCN gene
- Folate receptor
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- Author Affiliations
- 1. Department of Pediatrics, Chinese PLA General Hospital, 28 Fuxing Road, Beijing, 100853, China
- 2. Department of Hematology-Oncology, Capital Institute of Pediatrics, 2 Yabao Road, Beijing, 100020, China