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RISS 인기검색어
- 검색키워드
- 저자 : Lynne Hampson (검색결과 2 건)
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Identification of PlexinD1 and AHDC1 as a putative interactors for Tip-1 protein
Manal Aly Shalaby,Lynne Hampson,Anthony Oliver,Ian Hampson 한국유전학회 2011 Genes & Genomics Vol.33 No.4
The class 1 PDZ domain Tip-1 protein was first identified as a binding partner for the Human T-Cell Leukaemia Virus type 1 (HTLV-1) Tax oncoprotein. It was later shown to interact with: the RhoA signaling effector Rhotekin, the Wnt signaling effector β-catenin and the E6 oncoprotein from high-risk HPV16 but not low-risk HPV6. These observations suggested that Tip-1 may be an important “hub” protein that is involved in pathways with a proven link to carcinogenesis. Based on these findings, it was decided to further characterize the cellular role of Tip-1 by carrying out a yeast two-hybrid screen to identify new binding partners in order to uncover potentially novel functions of this protein. This identified an intracellular fragment of the trans-membrane receptor plexin D1 and a C-terminal fragment of the AT hook DNA binding containing 1 (AHDC1) protein which had a carboxyl terminal PDZ binding domain consensus sequence. Both of these interactions were confirmed by yeast mating assay which was also used to show that mutant constructs of AHDC1 lacking the carboxyl PDZ binding site did not bind Tip-1. Immunofluorescent imaging of these proteins in HPV16 E6 expressing human C33A cervical carcinoma cells suggested they may co-localize.
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Shin, Hye-Jin,Kim, Joo-Young,Hampson, Lynne,Pyo, Hongryull,Baek, Hye Jung,Roberts, Stephen A.,Hendry, Jolyon H.,Hampson, Ian N. Informa Healthcare 2010 International Journal of Radiation Biology Vol.86 No.9
<P><I>Purpose:</I> To examine the effect of the human papillomavirus (HPV) type 16-E6 (HPV ‘early’ gene) oncoprotein on in vitro radiosensitivity of HPV-negative/p53 mutant C33a cervical cancer cells.</P><P><I>Methods and materials:</I> The human cervical cancer cell line C33a was stably transfected with either the HPV16 E6 cDNA cloned into the vector pcDNA™3.0 (C33aE6) or the empty-vector control (C33aV). Radiosensitivity, DNA damage, and cell cycle measurements were made using standard clonogenic assays, immunofluorescent assessment of nuclear histone H2AX phosphorylated on serine-139 (γ-H2AX) foci, and flow cytometry. Western immunoblotting and fluorescence confocal microscopy were used to analyse the changes in cellular proteins. Real-time polymerase chain reaction (PCR) was used to compare levels of aurora A mRNA.</P><P><I>Results:</I> Compared to C33aV cells, C33aE6 cells showed enhanced radiation cell killing. This was associated with a large amount of polyploidy which was followed by late cell death in C33aE6 cells. Aurora A was highly expressed in C33aE6 cells at pre- and post-irradiation times compared to C33aV cells. Silencing aurora A resulted in a reduced amount of residual γ-H2AX foci in C33aE6 cells, and diminished the difference in radiosensitivity between the C33aE6 and C33aV cells.</P><P><I>Conclusion:</I> Our in vitro results indicate that genetic instability could be augmented in the HPV-infected cancer cells by up-regulation of aurora A, especially against a background of dysfunctional p53. Further studies are needed to examine whether aurora A could be a viable therapeutic target in HPV-related tumours.</P>
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