Abstract:
:Both transmission electron microscopy (TEM) and fluorescence spectrometry were used to reveal the characteristics of both subunit disassociation and recombination in apo-pig pancreas ferritin (apoPPF) in an alkaline medium ranging reversibly from pH 7.0 to 13.0. The experimental results indicated that apoPPF could be completely disassociated into 24 free subunits at pH 13.0, and then these subunits could be quickly reassembled into the original apoPPF once the pH of the reactive medium was returned to pH7.0. This novel and simple method could be used to effectively construct a novel nanometer cisplatin core-PPF (NCC-PPF). The major characteristics of NCC-PPF were investigated using various analytical methods such as ultraviolet-spectrometry, circular dichroism spectrometry and TEM, which indicated that its molecular structure was still similar to that of the original apoPPF. Results from the inductively coupled plasma mass spectrometer (ICP-MS) method showed that 11.26 cisplatin (CDDP) molecules were successfully packaged within the NCC-PPF shell, indicating that each molecule of apoPPF had the ability to enwrap 11.26 CDDP molecules for constructing the NCC-PPF. Flow cytometry showed that NCC-PPF also had the ability to release CDDP for inducing the apoptosis of gastric cancer cells BGC823 (GCC), but this phenomenon could scarcely be observed using apoPPF. A differential proteomic technique using two-dimensional electrophoresis (2-DE) gels selected and identified the differential proteins from cell apoptosis in order to reveal the molecular pathway of GCC apoptosis by both NCC-PPF and free CDDP, giving 13 differential expression proteins. These differential proteins could be further classified into six groups, which were described as being involved in the regulation of apoptosis, RNA transcription, oxidative stress response, signal transduction, cell metabolism, and cytoskeleton changes. In addition, a real-time PCR method was used to prove the expression level of mRNA and to identify the reliability of the protein expression according to these differential proteins, which indicated that the mRNA level changes of six differential proteins corresponded to those of its differential protein expression in 2-DE gels. These studies played an important role in reasonably revealing the different pathways of GCC apoptosis induced with both the CDDP released by NCC-PPF and the free CDDP. We thus suggest that apoPPF has great potential for constructing a nanometer carrier filled with various drugs for application in clinical work.
journal_name
J Proteomicsjournal_title
Journal of proteomicsauthors
Ji XT,Huang L,Huang HQdoi
10.1016/j.jprot.2012.03.013subject
Has Abstractpub_date
2012-06-18 00:00:00pages
3145-57issue
11eissn
1874-3919issn
1876-7737pii
S1874-3919(12)00146-7journal_volume
75pub_type
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pub_type: 临床试验,杂志文章
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journal_title:Journal of proteomics
pub_type: 杂志文章,评审
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journal_title:Journal of proteomics
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pub_type: 已发布勘误
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pub_type: 杂志文章
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journal_title:Journal of proteomics
pub_type: 临床试验,杂志文章
doi:10.1016/j.jprot.2012.06.004
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journal_title:Journal of proteomics
pub_type: 杂志文章
doi:10.1016/j.jprot.2016.07.029
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journal_title:Journal of proteomics
pub_type: 杂志文章
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journal_title:Journal of proteomics
pub_type: 杂志文章,评审
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journal_title:Journal of proteomics
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journal_title:Journal of proteomics
pub_type: 杂志文章
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journal_title:Journal of proteomics
pub_type: 杂志文章
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更新日期:2015-04-24 00:00:00
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journal_title:Journal of proteomics
pub_type: 杂志文章
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journal_title:Journal of proteomics
pub_type: 杂志文章
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更新日期:2016-05-17 00:00:00
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journal_title:Journal of proteomics
pub_type: 杂志文章
doi:10.1016/j.jprot.2018.01.002
更新日期:2018-03-20 00:00:00