1. Construction of HCC-targeting artificial miRNAs using natural miRNA precursors
- Author
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Zhenyu Jia and Xiaoming Huang
- Subjects
Cancer Research ,Oncogene ,Genetic enhancement ,Cell ,Articles ,hepatocellular carcinoma ,General Medicine ,Cell cycle ,Biology ,Bioinformatics ,microRNA precursor structure ,gene therapy ,digestive system diseases ,medicine.anatomical_structure ,Immunology and Microbiology (miscellaneous) ,artificial microRNA ,Cancer cell ,microRNA ,Cancer research ,medicine ,Luciferase ,Gene - Abstract
Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide, particularly in developing countries. Despite the achievements in clinical therapeutics, the HCC mortality rate remains high. A number of artificial microRNA (amiRNA)-based HCC gene therapy studies have demonstrated significant inhibition of invasion and induction of apoptosis of HCC cancer cells, indicating that this type of therapy may be a promising alternative to current therapeutics. Since the structure of the amiRNA precursor in the specific intracellular environment is critical for the processing to mature amiRNA, a precursor structure that may be efficiently processed is desired. In this study, we constructed amiRNAs targeting firefly luciferase with the precursor structures of six HCC-abundant microRNAs: miR-18a, miR-21, miR-192, miR-221, miR-222 and miR-224, and evaluated the processing efficiency of these amiRNAs in the HCC cell lines Hep3B and HepG2 using a luciferase reporter system. The results demonstrated that these amiRNA precursors are capable of being expressed in HCC cells, with the miR-221 precursor-based amiRNA exhibiting the most efficient inhibition on firefly luciferase at the levels of mRNA and protein activity. This finding provides a basis for constructing HCC-targeting amiRNAs with potent processing efficiency using the precursor structure of miR-221.
- Published
- 2013