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Simple and efficient gene transfer into the nucleus would facilitate non-viralgene delivery. One promising method of non-viral gene delivery is to apply penetrationenhancers. Chemicals, such as dimethyl sulfoxide (DMSO) and menthol, may havepromise as non-toxic vehicles in improving gene transfer efficiency. In this study,the cytotoxic effects of DMSO and menthol were evaluated using MTT assays. Genedelivery efficiency in a human breast cancer cell line (Bcap-37) was investigatedby quantitative PCR, fluorescence microscopy and flow cytometry. Non-toxic concentrationsof DMSO (2%) and menthol (12.5 µM) enhanced the efficiency of liposome-mediatedgene delivery in Bcap-37 cells. Quantitative PCR results showed that growth hormone(GH) mRNA expression in the post-menthol and pre-DMSO treatment groups was 10-foldhigher compared to that in the liposome group, while in the pre-menthol and post-DMSOtreatment groups, a 30-fold increase in GH mRNA expression was observed. BothDMSO and menthol treatments increased green fluorescent protein (GFP) expressionefficiency as shown by fluorescence microscopy experiments. Compared to the liposomegroup, the number of positive cells in the pre-menthol and post-DMSO treatmentgroups was significantly increased by 15%. Furthermore, cell cycle analysis demonstratedthat there were significant differences among the DMSO-treated group, the menthol-treatedgroup and the normal group, which implied different effects of DMSO and mentholtreatments. In conclusion, both non-toxic and harmless DMSO (2%) and menthol (12.5 µM)treatments improve gene transfer efficiency, while post-DMSO treatment may bethe most effective protocol in increasing transgene expression efficiency.

Enhancement of gene transfer efficiency in the Bcap-37 cell line by dimethyl sulphoxide and menthol