Summary
Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, prokaryotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS transformed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2+-resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentrations in 2 h. PEP-1-P27mt of 1 μmol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological protocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells.
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References
Lindgren M, Hallbrink M, Prochiantz A et al. Cell-penetrating peptides. Trends Pharmacol Sci, 2000,1(3):9–103
Morris M C, Depollier J, Mery J et al. A peptide carrier for the delivery of biologically active proteins into mammalian cells. Nat Biotechnol, 2001,19(12): 1173–1176
Polyak K, Kato J Y, Solomon M J et al. p27Kipl, a cyclin-Cdk inhibitor, links transforming growth factor-beta and contact inhibition to cell cycle arrest. Genes Dev, 1994,8(1):9–22
Polyak K, Lee M H, Erdjument-Bromage H et al. Cloning of p27Kip1, a cyclin-dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals. Cell, 1994,78(1):59–66
Park K H, Seol J Y. An adenovirus expressing mutant p27 showed more potent anti-tumor effects than adenovirus-p27 wildtype. Cancer Res, 2001,61:6163–6169
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YAN Shirong, female, born in 1965, Ph.D.
This project was supported by a grant from Department of Education of Hubei Province, China (No. Q200524001).
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Yan, S., Zhu, M., Qiu, F. et al. Study on the penetrability of PEP-1-P27mt for cell membranes in Vitro . J. Huazhong Univ. Sc. Technol. 27, 225–229 (2007). https://doi.org/10.1007/s11596-007-0302-2
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DOI: https://doi.org/10.1007/s11596-007-0302-2