New paper from Dang et al using Magnetofectamine Kit for miRNA transfection into HepB3, HepG2 and SNU449 hepatocellular carcinoma cell lines

200 nmol/L of miRNA were transfected in hepatocellular carcinoma cell lines (HepB3, HepG2 and SNU449) cultivated in 96-well plate, for 96 h using the Magnetofectamine transfection Kit from OZ Biosciences.

This paper demonstrated the high efficiency of Magnetofectamine kit from OZ Biosciences to transfect any kind of hepatocellular cell lines.

article reference: Asian Pac J Cancer Prev. 2014;15(12):4969-76.

Effects of miR-152 on Cell Growth Inhibition, Motility Suppression and Apoptosis Induction in Hepatocellular Carcinoma Cells.

Dang YW, Zeng J, He RQ, Rong MH, Luo DZ, Chen G.

Abstract

Background: miR-152 is involved in the genesis and development of several malignancies. However, its role in HCC has not been fully clarified. The aim of this study was to investigate the clinicopathological significance of miR-152 and its effect on the malignant phenotype of HCC cells. Methods: miR-152 expression was detected using real-time quantitative RT-PCR in 89 pairs of HCC formalin-fixed paraffin-embedded and their adjacent tissues. Functionally, in vitro effects and mechanisms of action of miR-152 on proliferation, viability, caspase activity, apoptosis and motility were explored in HepG2, HepB3 and SNU449 cells, as assessed by spectrophotometry, fluorimetry, fluorescence microscopy, wound-healing and Western blotting, respectively. Results: miR-152 expression in HCC was downregulated remarkably compared to that in adjacent hepatic tissues. miR-152 levels in groups of advanced clinical stage, larger tumor size and positive HBV infection, were significantly lower than in other groups. A miR-152 mimic could suppress cell growth, inhibit cell motility and increase caspase activity and apoptosis in HCC cell lines. Furthermore, Western blotting showed that the miR-152 mimic downregulated Wnt-1, DNMT1, ERK1/2, AKT and TNFRS6B signaling. Intriguingly, inverse correlation of TNFRF6B and miR-152 expression was found in HCC and bioinformatics confirmed that TNFRF6B might be a target of miR- 152. Conclusions: Underexpression of miR-152 plays a vital role in hepatocarcinogenesis and lack of miR-152 is related to the progression of HCC through deregulation of cell proliferation, motility and apoptosis. miR-152 may act as a tumor suppressor miRNA by also targeting TNFRSF6B and is therefore a potential candidate biomarker for HCC diagnosis, prognosis and molecular therapy.

Magnetofectamine from OZ Biosciences is a very efficient transfection reagent based on Magnetofection™ technology.