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Prenyl Ammonium Salts--New Carriers for Gene Delivery: A B16-F10 Mouse Melanoma Model.

Grecka E, Statkiewicz M, Gorska A, Biernacka M, Grygorowicz MA, Masnyk M, Chmielewski M, Gawarecka K, Chojnacki T, Swiezewska E, Malecki M - PLoS ONE (2016)

Bottom Line: AP-15/DOPE complexes were also efficient to introduce pDNA to cells, without much effect on cell viability.Furthermore, complexes containing AP-15 and therapeutic plasmid, encoding the TIMP metallopeptidase inhibitor 2 (TIMP2), introduced the TIMP2 gene with high efficiency to B16-F10 melanoma cells but not to B16-F10 melanoma tumors in C57BL/6 mice, as confirmed by TIMP2 protein level determination.Obtained results indicate that APs have a potential as non-viral vectors for cell transfection.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.

ABSTRACT

Purpose: Prenyl ammonium iodides (Amino-Prenols, APs), semi-synthetic polyprenol derivatives were studied as prospective novel gene transfer agents.

Methods: AP-7, -8, -11 and -15 (aminoprenols composed of 7, 8, 11 or 15 isoprene units, respectively) were examined for their capacity to form complexes with pDNA, for cytotoxicity and ability to transfect genes to cells.

Results: All the carriers were able to complex DNA. The highest, comparable to commercial reagents, transfection efficiency was observed for AP-15. Simultaneously, AP-15 exhibited the lowest negative impact on cell viability and proliferation--considerably lower than that of commercial agents. AP-15/DOPE complexes were also efficient to introduce pDNA to cells, without much effect on cell viability. Transfection with AP-15/DOPE complexes influenced the expression of a very few among 44 tested genes involved in cellular lipid metabolism. Furthermore, complexes containing AP-15 and therapeutic plasmid, encoding the TIMP metallopeptidase inhibitor 2 (TIMP2), introduced the TIMP2 gene with high efficiency to B16-F10 melanoma cells but not to B16-F10 melanoma tumors in C57BL/6 mice, as confirmed by TIMP2 protein level determination.

Conclusion: Obtained results indicate that APs have a potential as non-viral vectors for cell transfection.

No MeSH data available.


Related in: MedlinePlus

(a) cell viability and (b) total cell number of B16-F10 cells transfected with AP:pGFP complexes at various N/P ratios (r = 1.7 for AP-7, AP-8, AP-11, r = 2.0 for AP-15), or with AP-15/DOPE:pGFP and AP-15/DOPE/DMEM:pGFP lipoplexes at 2.5 μg of lipids/μg of pDNA dose, analysed by FACS.Transfections were performed with the use of 4 μg of pDNA. *P<0.05, **P<0.001.
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pone.0153633.g004: (a) cell viability and (b) total cell number of B16-F10 cells transfected with AP:pGFP complexes at various N/P ratios (r = 1.7 for AP-7, AP-8, AP-11, r = 2.0 for AP-15), or with AP-15/DOPE:pGFP and AP-15/DOPE/DMEM:pGFP lipoplexes at 2.5 μg of lipids/μg of pDNA dose, analysed by FACS.Transfections were performed with the use of 4 μg of pDNA. *P<0.05, **P<0.001.

Mentions: Furthermore, microscopic analysis revealed that transfection efficiency of AP-15/DOPE:pGFP complexes was substantially higher than the efficiency of Lipofectamine3000 (S2a and S2c Fig). Surprisingly, the results of flow cytometry indicated that Lipofectamine3000 transfected the cells more efficiently than AP-15/DOPE/DMEM or AP-15/DOPE transfection mixtures (Fig 3a). It should be kept in mind, however, that percentage of GFP-positive cells is dependent on the total number of the cells in the sample. In case of Lipofectamine3000 transfected cells the total cell count was considerably lower than that of cells transfected with AP-15 and DOPE-containing lipoplexes as indicated by flow cytometry (Fig 4b). Thus, our results suggest that Lipofectamine3000 significantly inhibited cell proliferation.


Prenyl Ammonium Salts--New Carriers for Gene Delivery: A B16-F10 Mouse Melanoma Model.

Grecka E, Statkiewicz M, Gorska A, Biernacka M, Grygorowicz MA, Masnyk M, Chmielewski M, Gawarecka K, Chojnacki T, Swiezewska E, Malecki M - PLoS ONE (2016)

(a) cell viability and (b) total cell number of B16-F10 cells transfected with AP:pGFP complexes at various N/P ratios (r = 1.7 for AP-7, AP-8, AP-11, r = 2.0 for AP-15), or with AP-15/DOPE:pGFP and AP-15/DOPE/DMEM:pGFP lipoplexes at 2.5 μg of lipids/μg of pDNA dose, analysed by FACS.Transfections were performed with the use of 4 μg of pDNA. *P<0.05, **P<0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835110&req=5

pone.0153633.g004: (a) cell viability and (b) total cell number of B16-F10 cells transfected with AP:pGFP complexes at various N/P ratios (r = 1.7 for AP-7, AP-8, AP-11, r = 2.0 for AP-15), or with AP-15/DOPE:pGFP and AP-15/DOPE/DMEM:pGFP lipoplexes at 2.5 μg of lipids/μg of pDNA dose, analysed by FACS.Transfections were performed with the use of 4 μg of pDNA. *P<0.05, **P<0.001.
Mentions: Furthermore, microscopic analysis revealed that transfection efficiency of AP-15/DOPE:pGFP complexes was substantially higher than the efficiency of Lipofectamine3000 (S2a and S2c Fig). Surprisingly, the results of flow cytometry indicated that Lipofectamine3000 transfected the cells more efficiently than AP-15/DOPE/DMEM or AP-15/DOPE transfection mixtures (Fig 3a). It should be kept in mind, however, that percentage of GFP-positive cells is dependent on the total number of the cells in the sample. In case of Lipofectamine3000 transfected cells the total cell count was considerably lower than that of cells transfected with AP-15 and DOPE-containing lipoplexes as indicated by flow cytometry (Fig 4b). Thus, our results suggest that Lipofectamine3000 significantly inhibited cell proliferation.

Bottom Line: AP-15/DOPE complexes were also efficient to introduce pDNA to cells, without much effect on cell viability.Furthermore, complexes containing AP-15 and therapeutic plasmid, encoding the TIMP metallopeptidase inhibitor 2 (TIMP2), introduced the TIMP2 gene with high efficiency to B16-F10 melanoma cells but not to B16-F10 melanoma tumors in C57BL/6 mice, as confirmed by TIMP2 protein level determination.Obtained results indicate that APs have a potential as non-viral vectors for cell transfection.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.

ABSTRACT

Purpose: Prenyl ammonium iodides (Amino-Prenols, APs), semi-synthetic polyprenol derivatives were studied as prospective novel gene transfer agents.

Methods: AP-7, -8, -11 and -15 (aminoprenols composed of 7, 8, 11 or 15 isoprene units, respectively) were examined for their capacity to form complexes with pDNA, for cytotoxicity and ability to transfect genes to cells.

Results: All the carriers were able to complex DNA. The highest, comparable to commercial reagents, transfection efficiency was observed for AP-15. Simultaneously, AP-15 exhibited the lowest negative impact on cell viability and proliferation--considerably lower than that of commercial agents. AP-15/DOPE complexes were also efficient to introduce pDNA to cells, without much effect on cell viability. Transfection with AP-15/DOPE complexes influenced the expression of a very few among 44 tested genes involved in cellular lipid metabolism. Furthermore, complexes containing AP-15 and therapeutic plasmid, encoding the TIMP metallopeptidase inhibitor 2 (TIMP2), introduced the TIMP2 gene with high efficiency to B16-F10 melanoma cells but not to B16-F10 melanoma tumors in C57BL/6 mice, as confirmed by TIMP2 protein level determination.

Conclusion: Obtained results indicate that APs have a potential as non-viral vectors for cell transfection.

No MeSH data available.


Related in: MedlinePlus