HYDROGEN-BOND SURROGATE PEPTIDES AND PEPTIDOMIMETICS FOR p53 REACTIVATION
a technology of p53 and surrogate peptides, which is applied in the direction of peptides, drug compositions, peptides, etc., can solve the problems of inability to recommend hpv vaccination for adults, poor vaccine uptake at 2%, and no clinical evidence to support, etc., to promote a pleiotropic, anti-cancer effect, and dramatic potentiation of the efficacy of cis-platinum
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example 1
Materials and Methods: Cell Lines
[0229]UMSCC47 and UMSCC74A were obtained from Dr. Thomas Carey at the University of Michigan. UPCI:SCC090 was provided by Dr. Susanne Gollin at the University of Pittsburgh (White et al., “The Influence of Clinical and Demographic Risk Factors on the Establishment of Head and Neck Squamous Cell Carcinoma Cell Lines,”Oral Oncol. 43(7):701-12 (2007), which is hereby incorporated by reference in its entirety). UMSCC47, UMSCC74A, and UPCI:SCC090 cells were grown in DMEM containing 10% FBS, 2 mM glutamine, 100 mg / mL streptomycin and 100 U / mL penicillin and maintained in a humidified atmosphere of 5% CO2 at 37° C.
example 2
Materials and Methods: Plasmid Construction and Transfection
[0230]The CH1 domain of p300 (nucleotides 332-417) was amplified by PCR (forward primer: 5′-GGATCCATGCCAGAGAAGCGCAAGCTCATCCAGC-3′; reverse primer: 5′-CTCGAGATCACCAGCATTTTTGAGGGGGAGACAC-3′) and inserted into pcDNA3.1 between the BamHI and Xhol restriction enzyme sites. UMSCC47, UMUPCI:SCC090, and UMSCC74A cells were transfected with pcDNA3.1 / empty or pcDNA3.1 / CH1 using Lipofectamine2000 (Invitrogen, Carlsbad, Calif.). Stable polyclonal populations were selected and maintained in the presence of G418 (Invitrogen).
example 3
Materials and Methods: Western Blot
[0231]Whole cell lysates were mixed with Laemmli loading buffer, boiled, separated by SDS-PAGE, and transferred to a nitrocellulose membrane. Subsequently, immunoblot analyses were performed using antibodies specific to V5 (Invitrogen), p53 (sc-126, Santa Cruz Biotechnology, Santa Cruz, Calif.), or acetylated[K382]-p53 (2525, Cell Signaling Technology). The signal was developed using the SuperSignal Western Blotting Kit (Pierce, Rockford, Ill.).
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