RESUMEN
The application of peptide drugs in cancer therapy is impeded by their poor biostability and weak cell permeability. Therefore, it is imperative to find biostable and cell-permeable peptide drugs for cancer treatment. Here, we identified a potent, selective, biostable, and cell-permeable cyclic d-peptide, NKTP-3, that targets NRP1 and KRASG12D using structure-based virtual screening. NKTP-3 exhibited strong biostability and cellular uptake ability. Importantly, it significantly inhibited the growth of A427 cells with the KRASG12D mutation. Moreover, NKTP-3 showed strong antitumor activity against A427 cell-derived xenograft and KRASG12D-driven primary lung cancer models without obvious toxicity. This study demonstrates that the dual NRP1/KRASG12D-targeting cyclic d-peptide NKTP-3 may be used as a potential chemotherapeutic agent for KRASG12D-driven lung cancer treatment.
Asunto(s)
Antineoplásicos , Neoplasias Pulmonares , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Humanos , Neoplasias Pulmonares/patología , Mutación , Péptidos/genética , Proteínas Proto-Oncogénicas p21(ras)/genéticaRESUMEN
Enrichment and detection of circulating free nucleic acids in biological samples have gained great attention for disease diagnosis or prognostic evaluation. Nanoscale metal-organic frameworks (NMOFs) have been used for aptamer-based nucleic acid sensing. In this work, different NMOFs, including ZIF-8, MIL-88, MIL-100, MIL-101, as well as Eu-TDA and Tb-TDA [prepared by the coordination of 2,2'-thiodiacetic acid (TDA) and Eu3+ or Tb3+], were investigated in nucleic acid sensing by employing their aptamer adsorption ability and fluorescence quenching capacity for the labeled dyes. Two types of dye aptamer, FAM-labeled aptamer (FAM-Ap) and TexasRedaptamer (TexasRed-Ap) were designed, and their adsorption properties on NMOFs-were compared. It was found that the TexasRed-Ap can be well used for nucleic acid (miR-21) extraction and sensing by linking with a pH-responsive nucleotide chain (TexasRed-Ap-pH) or with an additional random chain ssDNA-1' (TexasRed-Ap-a). After interacted with the target miR-21 in biosamples, the TexasRed-dsDNA + NMOFs composites can be collected, and the formed TexasRed-dsDNA can be released by changing pH value or addition of ssDNA-1, which is matched with ssDNA-1'. A linear relationship from 0.1 to 200 pM for miR-21 detection was obtained. The results show that the NMOFs can be used as promising platforms for nucleic acid extraction and fluorescent sensing.
Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Estructuras Metalorgánicas , Adsorción , ADN de Cadena SimpleRESUMEN
We report a series of colorable zeolitic imidazolate framework (ZIF)-based nanomaterials prepared by encapsulating starches (amylopectin, dextrin, or amylose) or tannic acid in the frameworks of ZIFs and first applied them in colorimetric assay of microRNA/DNA by adding I2/KI or FeCl3 solutions as chromogenic reagents. We found that iodine molecules can lead to rapid degradation of the ZIF-8 framework, while ZIF-90 remains stable. Therefore, ZIF-90 was selected for encapsulating the starches or tannic acid, and then assembled with polyethylenimine (PEI) and aptamers of microRNA/DNA. After interacting with the target microRNA/DNA, the aptamers (Ap) move away from the surface of the prepared Ap-starch@ZIF-90 or Ap-tan@ZIF-90, and the I2/KI or FeCl3 solution is added into the system to interact the starches (amylopectin, dextrin, or amylose) or tannic acid to generate different colors. According to the absorbance spectra, good linear correlations between the logarithm of absorbance intensity and the concentration of microRNA (1-180 nM) can be observed, and the naked eye can distinguish the change from â¼60 to â¼180 nM with a concentration gradient of 20 nM. A similar colorimetric assay ability for pathogenic bacteria can also be realized by detecting the gene fragments IS200 and eaeA. The detection limits can be potentially optimized by changing the amount of adsorbed PEI and aptamers on the surface of Ap-starch@ZIF-90 (or Ap-tan@ZIF-90) nanoparticles. This method could be a promising alternative for simple and cost-effective assay of microRNA/DNA.
Asunto(s)
Colorimetría , ADN/análisis , Imidazoles/química , Estructuras Metalorgánicas/química , MicroARNs/análisis , Zeolitas/química , Color , Humanos , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
Fontan surgery is a widely used palliative procedure that significantly improves the survival period of patients with complex congenital heart disease (CHD). However, it does not decrease postoperative complication rate. Previous studies suggested that elevated mean pulmonary artery pressure (mPAP) and vascular resistance lead to decreased exercise tolerance and myocardial dysfunction. Therapy with endothelial receptor antagonists (Bosentan) has been demonstrated to improve the patients' prognosis. A double-blind, randomized controlled trial was performed to explore the efficacy of Bosentan in treating patients who underwent the Fontan procedure. Eligible participants were randomly divided into Bosentan group and control group. Liver function was tested at a local hospital and the results were reported to the phone inspector every month. If the results suggested abnormal liver function, treatment would be adjusted or terminated. All the participants finished the follow-up study, with no patients lost to follow-up. Unblinding after 2-year follow-up, no mortality was observed in either group. However, secondary end-points were found to be significantly different in the comparable groups. The cardiac function and 6-min walking distance in the Bosentan group were significantly superior to those in the control group (P=0.018 and P=0.027). Bosentan could improve New York Heart Association (NYHA) functional status and improve the results of the 6-min walking test (6MWT) in Fontan patients post-surgery, and no other benefits were observed. Furthermore, a primary meta-analysis study systematically reviewed all the similar clinical trails worldwide and concluded an overall NYHA class improvement in Fontan patients who received Bosentan treatments.
Asunto(s)
Procedimiento de Fontan , Cardiopatías Congénitas/tratamiento farmacológico , Cardiopatías Congénitas/cirugía , Hígado/efectos de los fármacos , Sulfonamidas/administración & dosificación , Adolescente , Bosentán , Niño , Método Doble Ciego , Femenino , Estudios de Seguimiento , Cardiopatías Congénitas/patología , Humanos , Hígado/patología , Masculino , Cuidados Paliativos , Pronóstico , Resultado del TratamientoRESUMEN
Small interfering RNAs (siRNAs) are valuable reagents for efficient gene silencing in a sequencespecific manner via the RNA interference (RNAi) pathway. The current synthetic siRNA structure consists of symmetrical duplexes of 1921 base pairs (bp) with 2 nucleotide (nt) 3' overhangs. In this study, we report that an asymmetric siRNA (asiRNA) consisting of 17 bp duplex region (17 bp asiRNA) exhibited potent activity in inhibiting bcl-2 gene expression and cancer cell proliferation in vitro. Importantly, this asiRNA structure significantly reduced offtarget silencing by the sense strand. To improve the stability of the 17 bp asiRNA, we synthesized a series of chemically modified 17 bp asiRNAs. Further experiments showed that in comparison with the 17 bp asiRNA, the 17 bp asiRNAM2, one of the modified 17 bp asiRNAs, exhibited a comparable gene silencing activity and an improved stability in vitro. Furthermore, the 17 bp asiRNAM2 with a proteolipid micelle delivery system can effectively suppress the growth of H22 and BGC 803 tumors in vivo. These results suggest that the chemically modified asiRNAs may have potential as an effective therapeutic approach for cancer gene therapy in the future.
Asunto(s)
Proliferación Celular , Silenciador del Gen , Neoplasias Experimentales/prevención & control , Proteínas Proto-Oncogénicas c-bcl-2/antagonistas & inhibidores , ARN Interferente Pequeño/genética , Animales , Apoptosis , Citometría de Flujo , Células HeLa , Humanos , Técnicas In Vitro , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos ICR , Ratones Desnudos , Neoplasias Experimentales/metabolismo , Neoplasias Experimentales/patología , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
The ets-family transcription factor ETV6 (TEL) has been shown to be the target of a large number of balanced chromosomal translocations in various hematological malignancies and in some soft tissue tumors. Furthermore, ETV6 is essential for hematopoietic stem cell function. We identified ETV6 interacting proteins using the yeast two hybrid system. One of these proteins is the HIV Tat interacting protein (TIP60), a histone acetyltransferase (HAT) containing the highly conserved MYST domain. TIP60 functions as a corepressor of ETV6 in reporter gene assays. Fluorescently tagged ETV6 and TIP60 colocalize in the nucleus and an increase in nuclear localization of ETV6 was seen when TIP60 was cotransfected. ETV6 interacts with TIP60 through a 63 amino acids region located in the central domain of ETV6 between the pointed and the ets domain. The ETV6 interacting region of TIP60 mapped to the C2HC zinc finger of the TIP60 MYST domain. The interaction of TIP60 with full length ETV6 required an intact acetyltransferase domain of TIP60. Interestingly, the MYST domains of MOZ and MORF were also able to interact with portions of ETV6. These observations suggest that MYST domain HATs regulate ETV6 transcriptional activity and may therefore play critical roles in leukemogenesis and possibly in normal hematopoietic development.
Asunto(s)
Histona Acetiltransferasas/metabolismo , Proteínas Proto-Oncogénicas c-ets/metabolismo , Proteínas Represoras/metabolismo , Secuencia de Aminoácidos , Animales , Redes Reguladoras de Genes , Células HeLa , Histona Acetiltransferasas/química , Humanos , Ácidos Hidroxámicos/farmacología , Lisina Acetiltransferasa 5 , Metaloproteinasa 3 de la Matriz/genética , Ratones , Datos de Secuencia Molecular , Células 3T3 NIH , Unión Proteica/efectos de los fármacos , Mapeo de Interacción de Proteínas , Estructura Terciaria de Proteína , Transporte de Proteínas/efectos de los fármacos , Alineación de Secuencia , Fracciones Subcelulares , Técnicas del Sistema de Dos Híbridos , Proteína ETS de Variante de Translocación 6RESUMEN
The c-MYC oncogene encodes a transcription factor, which is sufficient and necessary for the induction of cellular proliferation. However, the c-MYC protein is a relatively weak transactivator suggesting that it may have other functions. To identify protein interactors which may reveal new functions or represent regulators of c-MYC we systematically identified proteins associated with c-MYC in vivo using a proteomic approach. We combined tandem affinity purification (TAP) with the mass spectral multidimensional protein identification technology (MudPIT). Thereby, 221 c-MYC-associated proteins were identified. Among them were 17 previously known c-MYC-interactors. Selected new c-MYC-associated proteins (DBC-1, FBX29, KU70, MCM7, Mi2-beta/CHD4, RNA Pol II, RFC2, RFC3, SV40 Large T Antigen, TCP1alpha, U5-116kD, ZNF281) were confirmed independently. For association with MCM7, SV40 Large T Antigen and DBC-1 the functionally important MYC-box II region was required, whereas FBX29 and Mi2-beta interacted via MYC-box II and the BR-HLH-LZ motif. In addition, regulators of c-MYC activity were identified: ectopic expression of FBX29, an E3 ubiquitin ligase, decreased c-MYC protein levels and inhibited c-MYC transactivation, whereas knock-down of FBX29 elevated the concentration of c-MYC. Furthermore, sucrose gradient analysis demonstrated that c-MYC is present in numerous complexes with varying size and composition, which may accommodate the large number of new c-MYC-associated proteins identified here and mediate the diverse functions of c-MYC. Our results suggest that c-MYC, besides acting as a mitogenic transcription factor, regulates cellular proliferation by direct association with protein complexes involved in multiple synthetic processes required for cell division, as for example DNA-replication/repair and RNA-processing. Furthermore, this first comprehensive description of the c-MYC-associated sub-proteome will facilitate further studies aimed to elucidate the biology of c-MYC.