Zeinab Salah*, Eman M. Abd El Azeem, Hanan F. Youssef , Amira M. Gamal-Eldeen*, Abdel R. Farrag, Emad El-Meliegy, Bangly Soliman and Mahmoud Elhefnawi* Pages 342 - 354 ( 13 )
Background: MicroRNA modulation therapy has shown great promise to treat hepatocellular carcinoma (HCC), however Efficient tissue-specific and safe delivery remains a major challenge.Objective: We sought to develop an inorganic-organic hybrid vehicle for the systemic delivery of the tumor suppressor miR-34a, and to investigate the efficiency of the delivered miR-34a in the treatment of HCC in vitro and in vivo. Methods: In the present study, pEGP-miR cloning and expression vector, expressing miR-34a, was electrostatically bound to polyethyleneimine (PEI), and then loaded onto ZSM-5 zeolite nanoparticles (ZNP). Qualitative and quantitative assessment of the transfection efficiency of miR-34a construct in HepG2 cells was applied by GFP screening and qRT-PCR, respectively. The expression of miR-34a target genes was investigated by qRT-PCR in vitro and in vivo. Results: ZNP/PEI/miR-34a nano-formulation could efficiently deliver into HepG2 cells with low cytotoxicity, indicating good biocompatibility of generated nanozeolite. Furthermore, five injected doses of ZNP/PEI/miR-34a nano-formulation in HCC induced male Balb-c mice, significantly inhibited tumor growth, and demonstrated improved cell structure, in addition to a significant decrease in alphafetoprotein level and liver enzymes activities, as compared to the positive control group. Moreover, injected ZNP/PEI/miR-34a nano-formulation led to a noticeable decrease in the CD44 and c-Myc levels. Results also showed that ZNP/PEI/miR-34a nano-formulation inhibited several target oncogenes including AEG-1, and SOX-9, in vitro and in vivo. Conclusion: Our results suggested that miR-34a is a powerful candidate in HCC treatment and that AEG-1 and SOX-9 are novel oncotargets of miR-34a in HCC. Results also demonstrated that our nano-formulation may serve as a candidate approach for miR-34a restoration for HCC therapy, and generally for safe gene delivery.
Hepatocellular carcinoma, tumor suppressor miRNA, replacement therapy, nanozeolite, gene delivery, oncotargets.
Biomedical Informatics and Chemo-Informatics Group Leader, Centre of Excellence for Medical Research, Informatics and System Dept, National Research Centre (NRC), Cairo, Department of Biochemistry, Faculty of Science, Ain Shams University, Cairo, Refractories, Ceramics and Building Materials Department, National Research Centre (NRC), Dokki, Cairo 12622, Clinical Laboratory Sciences Department, College of Applied Medical Sciences, Taif University, Al Mutamarat Rd, Al Mathnah, At Taif 26521, KSA / Cancer Biology and Genetics Laboratory, Centre of Excellence for Advanced Sciences, NRC, Cairo, Egypt / Biochemistry Department, NRC, Dokki, Cairo, Pathology Department Medical Division Research, National Research Centre, Cairo, 12622, Dokki, Department of Biomaterials, National Research Centre, Cairo, Biomedical Informatics and Chemo-Informatics Group Leader, Centre of Excellence for Medical Research, Informatics and System Dept, National Research Centre (NRC), Cairo, Biomedical Informatics and Chemo-Informatics Group Leader, Centre of Excellence for Medical Research, Informatics and System Dept, National Research Centre (NRC), Cairo