Gastric cancer is the second leading cause of cancer-related death worldwide. RNA nanotechnology has recently emerged as an important field due to recent finding of its high thermodynamic stability, favorable and distinctive in vivo attributes. Here we reported the use of the thermostable three-way junction (3WJ) of bacteriophage phi29 motor pRNA to escort folic acid, a fluorescent image marker and BRCAA1 siRNA for targeting, imaging, delivery, gene silencing and regression of gastric cancer in animal models. In vitro assay revealed that the RNA nanoparticles specifically bind to gastric cancer cells, and knock-down the BRCAA1 gene. Apoptosis of gastric cancer cells was observed. Animal trials confirmed that these RNA nanoparticles could be used to image gastric cancer in vivo, while showing little accumulation in crucial organs and tissues. The volume of gastric tumors noticeably decreased during the course of treatment. No damage to important organs by RNA nanoparticles was detectible. All the results indicated that this novel RNA nanotechnology can overcome conventional cancer therapeutic limitations and opens new opportunities for specific delivery of therapeutics to stomach cancer without damaging normal cells and tissues, reduce the toxicity and side effect, improve the therapeutic effect, and exhibit great potential in clinical tumor therapy.
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This work is supported by Chinese Key Basic Research Program (973 Project) (No. 2010CB933901 and 2015CB931802), the National Natural Scientific Foundation of China (Grant No. 81225010, 81327002, and 31170961) and NIH grants CA151648 (P.G.), EB019036 (P.G.) and EB012135 (P.G.). Funding to Peixuan Guo’s Endowed Chair in Nanobiotechnology at University of Kentucky is by the William Farish Endowment Fund. AFM images were obtained at Nanoimaging Core Facility supported by NIH SIG program and University of Nebraska Medical Center Program of ENRI to Luda Shlyakhtenko and Yuri Lyubchenko.
Cui, Daxiang; Zhang, Chunlei; Liu, Bing; Shu, Yi; Du, Tong; Shu, Dan; Wang, Kan; Dai, Fangping; Liu, Yanlei; Li, Chao; Pan, Fei; Yang, Yuming; Ni, Jian; Li, Hui; Brand-Saberi, Beate; and Guo, Peixuan, "Regression of Gastric Cancer by Systemic Injection of RNA Nanoparticles Carrying Both Ligand and siRNA" (2015). Markey Cancer Center Faculty Publications. 36.
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Figure 1: Global structure of the therapeutic RNA nanoparticles with BRCAA1 siRNA.
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Figure 2: Flow cytometry analysis for specific binding of 3WJ-FA-A647 nanoparticles to MGC803 cells (left, folate positive), GES-1 cells (right, folate negative control).
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Figure 4: Inhibition of the growth of MGC803 cells by the nanoparticle of FA-pRNA-3WJ-BRCAA1siRNA using CCK8 (Cell Counting Kit-8) assays.
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Figure 5: Determination of cell death by flow cytometry of Annexin V-FITC/PI staining in MGC803 cells transfected with 25 nM FA-pRNA-3WJ-BRCAA1siRNA or FA-pRNA-3WJ-Scram-siRNA for 48 h.
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Figure 8: Tumor size curve as the post-treatment time increases.
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Figure 9: Result of HE immunostaining of important organs showing the undetectable damage.
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Figure 10: The expression of related apoptosis proteins of MGC803 cells at 48 h post-treatment by Western blotting.
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Figure 11: The potential mechanism of RNA nanoparticles for gastric cancer therapy.