Mechanisms of Cellular Uptake with Chitosan/DNA Complex in Hepatoma Cell Line

Abstract:

Article Preview

Chitosan (CS) has a high potential for gene delivery into mammalian cells. However, its uptake mechanism is not well clarified. We investigated the effects of inhibitors of clathrin-mediated endocytosis (chlorpromazine), caveolae-mediated endocytosis (genistein), macropinocytosis (LY 29004 and wortmannin), microtubuli polymerization (nocodazole) and of membrane cholesterol recycle (methyl-β-cyclodextrin) on the transfection efficiency with CS/pEGFP complexes and on the internalization of CS/rhodamine-labeled pEGFP complexes by hepatoma cell line (Huh 7 cells). The transfection was blocked by nocodazole, genistein, and methyl-β-cyclodextrin, respectively. CS/DNA complexes internalization was clearly inhibited by genistein. We conclude that the complexes uptake predominantly by caveolin-mediated pathways. In addition, fluorescence colocalization studies with acidotropic probes, LysoSensor dye, illustrated that CS/DNA complexes are targeted to lysosomes for the degradation after internalization.

Info:

Periodical:

Edited by:

Tawee Tunkasiri

Pages:

485-488

DOI:

10.4028/www.scientific.net/AMR.506.485

Citation:

A. Apirakaramwong et al., "Mechanisms of Cellular Uptake with Chitosan/DNA Complex in Hepatoma Cell Line", Advanced Materials Research, Vol. 506, pp. 485-488, 2012

Online since:

April 2012

Export:

Price:

$35.00

[1] F.C. MacLaughlin, R.J. Mumper, et al.: J Control Release. Vol. 56 (1998), p.259.

[2] R. Jayakumar, K.P. Chennazhi, et al.: Carbohydr Polym. Vol. 79 (2010), 1.

[3] W.G. Liu, K.D. Yao.: J Control Release. Vol. 83 (2002), p.1.

[4] S.D. Conner, S.L. Schmid. Nature. Vol. 422 (2003), p.37.

[5] S. Huth, J. Lausier. et al.: J Gene Med. Vol. 6 (2004), p.923.

[6] W. Weecharangsan, P. Opanasopit, et al.: AAPS PharmSciTech. Vol. 7 (2006), p. E1.

[7] U.S. Huth, R. Schubert, R. Peschka-Süss.: J Control Release. Vol. 110 (2006), p.490.

[8] M.A. E.M. van der Aa, U.S. Huth, et al.: Pharm Res. Vol. 24 (2007), p.1590.

[9] M.D. Smith, J.C. Barbenel, et al.: Int J Artif Organs. Vol. 15 (1992), p.191.

[10] P. L. Leopold, G. Kreitzer. et al.: Hum. Gene Ther. Vol. 11 (2000), p.151.

In order to see related information, you need to Login.