RESUMEN
The ubiquitin-proteasome system (UPS) is essential for maintaining cell homeostasis by orchestrating the protein degradation, but is impaired in various diseases, including cancers. Several proteasome inhibitors, such as bortezomib, are currently used in cancer treatment, but associated toxicity limits their widespread application. Recently metal complex-based drugs have attracted great attention in tumor therapy; however, their application is hindered by low water-solubility and poor absorbency. Herein, we synthesized a new type of gold (I) complex named Na-AuPT, and further characterized its anticancer activity. Na-AuPT is highly water-soluble (6 mg/mL), and it was able to potently inhibit growth of a panel of 11 cancer cell lines (A549, SMMC7721, H460, HepG2, BEL7402, LNCap, PC3, MGC-803, SGC-7901, U266, and K562). In A549 and SMMC7721 cells, Na-AuPT (in a range of 2.5-20 µM) inhibited the UPS function in a dose-dependent fashion by targeting and inhibiting both 20 S proteasomal proteolytic peptidases and 19 S proteasomal deubiquitinases. Furthermore, Na-AuPT induced caspase-dependent apoptosis in A549 and SMMC7721 cells, which was prevented by the metal chelator EDTA. Administration of Na-AuPT (40 mg · kg-1 · d-1, ip) in nude mice bearing A549 or SMMC7721 xenografts significantly inhibited the tumor growth in vivo, accompanied by increased levels of total ubiquitinated proteins, cleaved caspase 3 and Bax protein in tumor tissue. Moreover, Na-AuPT induced cell death of primary mononuclear cells from 5 patients with acute myeloid leukemia ex vivo with an average IC50 value of 2.46 µM. We conclude that Na-AuPT is a novel metal-based proteasome inhibitor that may hold great potential for cancer therapy.
Asunto(s)
Antineoplásicos , Neoplasias , Animales , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Apoptosis , Línea Celular Tumoral , Humanos , Ratones , Ratones Desnudos , Neoplasias/tratamiento farmacológico , Complejo de la Endopetidasa Proteasomal/metabolismo , Inhibidores de Proteasoma/farmacología , Inhibidores de Proteasoma/uso terapéutico , Ubiquitina/metabolismo , AguaRESUMEN
A new neutral polysaccharide, named AGP, was extracted from glycyrrhiza residue by 5% NaOH alkaline solution and purified by DEAE-celluloseand Sephadex G-150. A single and symmetrical peak was shown by HPLC, indicating that AGP is a homogeneous polysaccharide with a molecular weight of 2.89 × 103 KDa. Thespecific rotation of AGP was detected by a polarimeter and it was +45°. Monosaccharide composition analysis indicated that AGP was consisted of l-rhamnose: l-arabinose: d-xylose: d-mannose: d-glucose and d-galactose with a molar ratio of 1:2.33:2.85:0.69:3.05:1.54. The structure of AGP was analyzed by GC-MS, periodate oxidation, Smith degradation, FT-IR, methylation and NMR, which indicated that the AGP was composed of â 6)-ß-d-Glcp-( â backbone and the â4)-α-d-Xylp-(1â, â5)-α-l-Araf-(1â, â3)-α-l-Rhap-(1â, â6)-α-d-Galp-(1â, â3,6)-α-Manp-(1â and â1)-ß-d-Glcp as branches. The results of Congo red experiment and circular dichroism (CD) showed that there was triple helix conformation in AGP. The micro-structure of AGP were detected by scanning electron microscopy (SEM), which concluded that the shape of AGP was a "thin slice" and its structure is not regular. The crystal configuration was identified by X-ray diffraction (XRD), showing that there is no crystal structure. Furthermore, the AGP exhibited certain inhibition activity on α-glucosidase.