Chitosan Combined with Molecular Beacon for Mir-155 Detection and Imaging in Lung Cancer

Abstract

Lung cancer is the major cause of cancer-related deaths worldwide, thus developing effective methods for its early diagnosis is urgently needed. In recent years, microRNAs (miRNAs, miR) have been reported to play important roles in carcinogenesis and have become potential biomarkers for cancer diagnosis and treatment. Molecular beacon (MB) technology is a universal technology to detect DNA/RNA expression in living cells. As a natural polymers, chitosan (CS) nanoparticles could be used as a carrier for safe delivery of nucleic acid. In this study, we developed a probe using nanoparticles of miR-155 MB self assembled with CS (CS-miR-155 MB) to image the expression of miR-155 in cancer cells. Hybridization assay showed that the locked nucleic acid (LAN) modified miR-155 MB could target miR-155 effectively and sensitively. The miR-155 MB self-assembly with CS nanoparticles formed stable complexes at the proper weight ratio. The CS nanoparticles showed higher fluorescence intensity and transfection efficiency than the lipid-based formulation transfection agent by confocal microscopy and flow cytometry analysis. The CS-MB complexes were found to be easily synthesized and exhibited strong OPEN ACCESSMolecules 2014, 19 14711 enzymatic stability, efficient cellular uptake, high target selectivity and biocompatibility. The CS-MB complexes can also be applied in other cancers just by simply changing for a targeted miRNA highly expressed in those cancer cells. Therefore, it is a promising vehicle used for detecting miRNA expression in living cells.

Detection of miR-155-5p and imaging lung cancer for early diagnosis: in vitro and in vivo study

“…Based on a previous study (Zhu et al 2014), CS nanoparticles (Guanghan Hengyu, Sichuan, China), hsa-miR-155-5p MB, mmu-miR-155-5p MB, or random sequence MB (RS MB) (Sangon, Shanghai, China) were mixed at the ratio of Wcs/W MB = 7:1 to synthesize the CS-MB nanoparticles. Hsa-miR-155-5p MB (5′-Cy5-C + CAGCG-ACC + CCT + ATCA + CGAT + TAG CAT TAA-CGCT + GG-BHQ3-3′) and Mmu-miR-155-5pMB (5′-Cy5-C + CAGCG -ACC + CCT + ATCA + CAAT + TAG CAT TAA-CGCT + GG-BHQ3-3′) were designed according to the miR-155-5p sequences.…”

“…Chitosan (CS) is a natural cationic polysaccharide and has been widely used to mediate the gene transfection of plasmids and small interfering RNAs (siRNAs) (Cao et al 2019;Rahmani et al 2015;Jaiswal et al 2019). In a previous study (Zhu et al 2014), we used CS nanoparticles combined with MB for the detection and imaging miR-155-5p in non-small cells lung cancer.…”

Identification and imaging of miR-155 in the early screening of lung cancer by targeted delivery of octreotide-conjugated chitosan-molecular beacon nanoparticles

“…However, the CS-RS MB group showed no red fluorescence signal but only the green fluorescence was emitted by the fluorescent dye FITC. By combining the previous fluorescence intensity analysis (Figure S2) (Zhu et al., 2014). The fluorescence intensity of PC-3, A549, and SPC-A1 cells appeared to be stronger after miR-155 MB transfection in CS-MB-OCT group than that of CS-MB transfection group and siPORT liposome transfection group, indicating that OCT can target and bind to SSTR2.…”

“…Chitosan (CS) nanoparticles had been widely used to mediate gene transfection such as plasmids, siRNAs, microRNA, and demonstrated high safety and effectiveness (Geng et al., 2013; Raftery et al., 2013). In the previous studies, we have successfully used chitosan nanoparticles as miR-155 molecular beacon vehicles to detect miR-155 and imaging in the lung cancer cells (Zhu et al., 2014).…”

The effects of chitosan/miR-200c nanoplexes on different stages of cancers in breast cancer cell lines

“…Chitosan-molecular beacon complexes were used for miR-155 detection and imaging in lung cancer (Zhu et al, 2014). In another paper, Chen et al (2014) used the Achilles tendon injured rat model and investigated the role of miRNAs on the effects of chitosan on tendon healing via the regulation of the TGF-β1/Smad3 pathway.…”