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

Lin,Jian; Zhu,Li Qi; Qin,Tao; Yu,Qing Hua; Yang,Qian

doi:10.3892/mmr.2012.1084

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

Authors:
- Jian Lin
- Li Qi Zhu
- Tao Qin
- Qing Hua Yu
- Qian Yang
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Affiliations: Key Laboratory of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China
Published online on: September 17, 2012 https://doi.org/10.3892/mmr.2012.1084
Pages: 1293-1300

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Abstract

Simple and efficient gene transfer into the nucleus would facilitate non-viral gene delivery. One promising method of non-viral gene delivery is to apply penetration enhancers. Chemicals, such as dimethyl sulfoxide (DMSO) and menthol, may have promise as non-toxic vehicles in improving gene transfer efficiency. In this study, the cytotoxic effects of DMSO and menthol were evaluated using MTT assays. Gene delivery efficiency in a human breast cancer cell line (Bcap-37) was investigated by quantitative PCR, fluorescence microscopy and flow cytometry. Non-toxic concentrations of DMSO (2%) and menthol (12.5 µM) enhanced the efficiency of liposome-mediated gene 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-fold higher compared to that in the liposome group, while in the pre-menthol and post-DMSO treatment groups, a 30-fold increase in GH mRNA expression was observed. Both DMSO and menthol treatments increased green fluorescent protein (GFP) expression efficiency as shown by fluorescence microscopy experiments. Compared to the liposome group, the number of positive cells in the pre-menthol and post-DMSO treatment groups was significantly increased by 15%. Furthermore, cell cycle analysis demonstrated that there were significant differences among the DMSO-treated group, the menthol-treated group and the normal group, which implied different effects of DMSO and menthol treatments. In conclusion, both non-toxic and harmless DMSO (2%) and menthol (12.5 µM) treatments improve gene transfer efficiency, while post-DMSO treatment may be the most effective protocol in increasing transgene expression efficiency.

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1	Lechardeur D and Lukacs GL: Intracellular barriers to non-viral gene transfer. Curr Gene Ther. 2:183–194. 2002. View Article : Google Scholar : PubMed/NCBI
2	Leong KW, Mao HQ, Truong-Le VL, Roy K, Walsh SM and August JT: DNA-polycation nanospheres as non-viral gene delivery vehicles. J Control Release. 53:183–193. 1998. View Article : Google Scholar : PubMed/NCBI
3	Kamiya H, Tsuchiya H, Yamazaki J and Harashima H: Intracellular trafficking and transgene expression of viral and non-viral gene vectors. Adv Drug Deliv Rev. 52:153–164. 2001. View Article : Google Scholar : PubMed/NCBI
4	Zhou M, Liu H, Xu X, et al: Identification of nuclear localization signal that governs nuclear import of BRD7 and its essential roles in inhibiting cell cycle progression. J Cell Biochem. 98:920–930. 2006. View Article : Google Scholar : PubMed/NCBI
5	Man N, Yu L, Zheng F, Li Y and Wen LP: Efficient gene transfer to rat fetal osteoblastic cells by synthetic peptide vector system. Protein Pept Lett. 16:368–372. 2009. View Article : Google Scholar : PubMed/NCBI
6	Notman R, den Otter WK, Noro MG, Briels WJ and Anwar J: The permeability enhancing mechanism of DMSO in ceramide bilayers simulated by molecular dynamics. Biophys J. 93:2056–2068. 2007. View Article : Google Scholar : PubMed/NCBI
7	Ribbeck K and Gorlich D: The permeability barrier of nuclear pore complexes appears to operate via hydrophobic exclusion. Embo J. 21:2664–2671. 2002. View Article : Google Scholar : PubMed/NCBI
8	Brunner S, Sauer T, Carotta S, Cotten M, Saltik M and Wagner E: Cell cycle dependence of gene transfer by lipoplex, polyplex and recombinant adenovirus. Gene Ther. 7:401–407. 2000. View Article : Google Scholar : PubMed/NCBI
9	Guillard EC, Laugel C and Baillet-Guffroy A: Molecular interactions of penetration enhancers within ceramides organization: a FTIR approach. Eur J Pharm Sci. 36:192–199. 2009. View Article : Google Scholar : PubMed/NCBI
10	Hewitt PG, Poblete N, Wester RC, Maibach HI and Shainhouse JZ: In vitro cutaneous disposition of a topical diclofenac lotion in human skin: effect of a multi-dose regimen. Pharm Res. 15:988–992. 1998. View Article : Google Scholar : PubMed/NCBI
11	Mittal A, Sara UV, Ali A and Aqil M: The effect of penetration enhancers on permeation kinetics of nitrendipine in two different skin models. Biol Pharm Bull. 31:1766–1772. 2008. View Article : Google Scholar : PubMed/NCBI
12	Li L, Shen W, Min L, Dong H, Sun Y and Pan Q: Human lactoferrin transgenic rabbits produced efficiently using dimethylsulfoxide-sperm-mediated gene transfer. Reprod Fertil Dev. 18:689–695. 2006. View Article : Google Scholar
13	Brain KR, Green DM, Dykes PJ, Marks R and Bola TS: The role of menthol in skin penetration from topical formulations of ibuprofen 5% in vivo. Skin Pharmacol Physiol. 19:17–21. 2006.PubMed/NCBI
14	Ho HO, Chen LC, Lin HM and Sheu MT: Penetration enhancement by menthol combined with a solubilization effect in a mixed solvent system. J Control Release. 51:301–311. 1998. View Article : Google Scholar : PubMed/NCBI
15	Bremner KH, Seymour LW, Logan A and Read ML: Factors influencing the ability of nuclear localization sequence peptides to enhance nonviral gene delivery. Bioconjug Chem. 15:152–161. 2004. View Article : Google Scholar : PubMed/NCBI
16	Jain PT and Gewirtz DA: Enhancement of liposomal gene delivery in human breast cancer cells by dimethyl sulfoxide. Int J Mol Med. 1:609–611. 1998.PubMed/NCBI
17	Fiore M, Zanier R and Degrassi F: Reversible G(1) arrest by dimethyl sulfoxide as a new method to synchronize Chinese hamster cells. Mutagenesis. 17:419–424. 2002. View Article : Google Scholar : PubMed/NCBI
18	Escriou V, Carriere M, Bussone F, Wils P and Scherman D: Critical assessment of the nuclear import of plasmid during cationic lipid-mediated gene transfer. J Gene Med. 3:179–187. 2001. View Article : Google Scholar : PubMed/NCBI
19	Mir LM: Nucleic acids electrotransfer-based gene therapy (electrogenetherapy): past, current, and future. Mol Biotechnol. 43:167–176. 2009. View Article : Google Scholar : PubMed/NCBI
20	Ramezani A and Hawley RG: Strategies to insulate lentiviral vector-expressed transgenes. Methods Mol Biol. 614:77–100. 2010. View Article : Google Scholar : PubMed/NCBI
21	Heckert RA, Elankumaran S, Oshop GL and Vakharia VN: A novel transcutaneous plasmid-dimethylsulfoxide delivery technique for avian nucleic acid immunization. Vet Immunol Immunopathol. 89:67–81. 2002. View Article : Google Scholar : PubMed/NCBI
22	Sultana Y, Jain R, Aqil M and Ali A: Review of ocular drug delivery. Curr Drug Deliv. 3:207–217. 2006. View Article : Google Scholar : PubMed/NCBI
23	Wang H, Zhong CY, Wu JF, Huang YB and Liu CB: Enhancement of TAT cell membrane penetration efficiency by dimethyl sulphoxide. J Control Release. 143:64–70. 2010. View Article : Google Scholar : PubMed/NCBI
24	Campeau P, Chapdelaine P, Seigneurin-Venin S, Massie B and Tremblay JP: Transfection of large plasmids in primary human myoblasts. Gene Ther. 8:1387–1394. 2001. View Article : Google Scholar : PubMed/NCBI
25	Migita S, Hanagata N, Tsuya D, Yamazaki T, Sugimoto Y and Ikoma T: Transfection efficiency for size-separated cells synchronized in cell cycle by microfluidic device. Biomed Microdevices. 13:725–729. 2011. View Article : Google Scholar : PubMed/NCBI
26	Srinivas S, Sironmani TA and Shanmugam G: Dimethyl sulfoxide inhibits the expression of early growth-response genes and arrests fibroblasts at quiescence. Exp Cell Res. 196:279–286. 1991. View Article : Google Scholar : PubMed/NCBI
27	Collas P and Alestrom P: Rapid targeting of plasmid DNA to zebrafish embryo nuclei by the nuclear localization signal of SV40 T antigen. Mol Mar Biol Biotechnol. 6:48–58. 1997.PubMed/NCBI
28	Arenal A, Pimentel R, Garcia C, Pimentel E and Alestrom P: The SV40 T antigen nuclear localization sequence enhances nuclear import of vector DNA in embryos of a crustacean (Litopenaeus schmitti). Gene. 337:71–77. 2004. View Article : Google Scholar : PubMed/NCBI
29	Medina-Kauwe LK, Xie J and Hamm-Alvarez S: Intracellular trafficking of nonviral vectors. Gene Ther. 12:1734–1751. 2005. View Article : Google Scholar : PubMed/NCBI
30	Xavier J, Singh S, Dean DA, Rao NM and Gopal V: Designed multi-domain protein as a carrier of nucleic acids into cells. J Control Release. 133:154–160. 2009. View Article : Google Scholar : PubMed/NCBI
31	Dean DA, Strong DD and Zimmer WE: Nuclear entry of nonviral vectors. Gene Ther. 12:881–890. 2005. View Article : Google Scholar : PubMed/NCBI