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INTRODUCTION: Over the past decade, we have moved on from a predominantly morphological and clinical classification of myeloproliferative neoplasms (MPN) to a more evolved classification that accounts for the molecular heterogeneity that is unique to this subgroup of hematological malignancies. This usually incorporates mutations in Janus kinase 2 (JAK2), MPL, and calreticulin (CALR) genes. In this manuscript, we report the frequency of these mutations in a cohort of Indian patients at a tertiary cancer center. MATERIALS AND METHODS: One hundred and thirty cases of MPN were included in this study. These cases were diagnosed and classified based on the World Health Organization 2008 criteria. JAK2 and MPL mutations were detected using high sensitivity allele-specific polymerase chain reaction using fluorescent labeled primers followed by capillary electrophoresis. A subset of JAK2 and CALR mutations were assessed using a fragment length assay. RESULTS: Among the MPN, we had 20 cases of polycythemia vera (PV), 34 cases of essential thrombocythemia (ET), and 59 of myelofibrosis (MF). JAK2, MPL, and CALR mutations were mutually exclusive of each other. Seventeen cases were categorized as MPN unclassifiable (MPN-U). JAK2p.V617F and MPL mutations were present in 60% (78 of 130) and 5.3% (7 of 130) of all MPN. All the PV cases harbored the JAK2 p.V617F mutation. A total of 23.8% (31 of 130) of patients harbored CALR mutations. CALR exon 9 mutations were detected in 60.8% (14 of 23) and 50% (5 of 10) of JAK2 and MPL negative MF and ET cases, respectively. MPN-U cases included three JAK2 p.V617F positive, two MPL p.W515 L, and 12 CALR positive cases. Ten different types of CALR indels (8 deletions and 2 insertions) were detected of which Type I and Type II mutations were the most common, occurring at a frequency of 45.1% (14 of 31) and 22.5% (7 of 31), respectively. DISCUSSION AND CONCLUSION: We report frequencies of JAK2 p. V617F, MPL exon 10 and CALR mutations in 130 patients similar to those reported in western literature. These mutations carry not only diagnostic but also prognostic relevance.
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Calreticulina/genética , Proteínas de Fusión bcr-abl/genética , Janus Quinasa 2/genética , Trastornos Mieloproliferativos/genética , Receptores de Trombopoyetina/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , India , Masculino , Persona de Mediana Edad , Mutación/genética , Policitemia Vera/genética , Mielofibrosis Primaria/genética , Estudios Retrospectivos , Trombocitemia Esencial/genética , Adulto JovenRESUMEN
Here, we report a non-invasive strategy for isolating cancer cells by autonomously propelled carbon nanotube (CNT) microrockets. H2O2-driven oxygen (O2) bubble-propelled microrockets were synthesized using CNT and Fe3O4 nanoparticles in the inner surface and covalently conjugating transferrin on the outer surface. Results show that self-propellant microrockets can specifically capture cancer cells.
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Nanotubos de Carbono/química , Células Neoplásicas Circulantes/química , Óxido Ferrosoférrico/química , Células HCT116 , Humanos , Peróxido de Hidrógeno/química , Ligandos , Nanopartículas de Magnetita/química , Microscopía Electrónica de Transmisión , Oxígeno/química , Propiedades de Superficie , Imagen de Lapso de TiempoRESUMEN
In a first series from India, we report 32 cases of lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (LPL/WM) over 7 years. Here, we analyzed 32 patients with LPL/WM for MYD88 L265P mutation and correlated mutation staus with hematological and biochemical parameters and also with the International Prognostic Scoring System (ISSWM) and treatment response. Twenty-seven out of 32 cases of LPL/WM (84.3%) harbored the MYD88 L265P mutation. MYD88 wild-type WM was associated with a lower number of tumor cells (p<0.01) and older age (p=0.02) and a lower ISSWM score at presentation (p=0.03) as compared to mutated LPL/WM. On evaluation of response (n=23), 44.4% of patients with MYD88 mutated LPL/WM had progressive disease, whereas no patient in the MYD88 unmutated group changed their baseline status. We confirm the high frequency of MYD88 mutations in LPL/WM. Although the number of MYD88 wild-type cases was limited, our data indicate that MYD88 may represent an adverse prognostic marker for LPL/WM.
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Mutación Missense , Factor 88 de Diferenciación Mieloide/genética , Macroglobulinemia de Waldenström/genética , Macroglobulinemia de Waldenström/patología , Anciano , Secuencia de Bases , Biomarcadores de Tumor/genética , Análisis Mutacional de ADN , Progresión de la Enfermedad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Pronóstico , Inducción de Remisión , Estudios Retrospectivos , Resultado del Tratamiento , Macroglobulinemia de Waldenström/tratamiento farmacológicoRESUMEN
Carbon nanostructures such as multiwalled carbon nanotubes (CNT) and graphene (G) are potential candidates in a large number of biomedical applications. However, there is limited understanding and connection between the physicochemical properties of diverse carbon nanostructures and biological systems, particularly with regard to cellular responses. It is also crucial to understand how the structure and surface composition of carbon nanostructures affect the cellular internalization process. Here, through in vitro cellular entry kinetics and cytotoxicity studies using MCF-7 breast cancer cells and H460 human lung cancer cells, we show that the structure and surface composition of CNT and G conjugates with various molecules such as PAMAM dendrimers (G4) and G4-poly(ethylene glycol) (PEG) are directly related to their cellular internalization ability and toxicity. Interestingly, the cellular association of CNT and G nanoconjugates was observed to be structure and surface composition dependent. We found that CNT conjugates internalized more compared to G conjugates. Furthermore, G4 conjugated CNT internalized more compared to G4-PEG conjugated CNT, whereas, higher internalization was found for G4-PEG conjugated G than G4 conjugated G. We have also correlated the cytotoxicity and cellular uptake mechanisms of CNT, G, and their conjugates through zeta potential measurements, fluorescence quenching studies and by fluorescence-activated cell sorting. Altogether these studies suggest different biological activities of the carbon nanostructures, with the shape and surface composition playing a primary role.
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A multicomponent magneto-dendritic nanosystem (MDNS) is designed for rapid tumor cell targeting, isolation, and high-resolution imaging by a facile bioconjugation approach. The highly efficient and rapid-acting MDNS provides a convenient platform for simultaneous isolation and high-resolution imaging of tumor cells, potentially leading towards an early diagnosis of cancer.
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Separación Celular/métodos , Separación Inmunomagnética/métodos , Técnicas de Diagnóstico Molecular/métodos , Nanopartículas , Células Neoplásicas Circulantes/metabolismo , Células Neoplásicas Circulantes/patología , Transferrina/farmacocinética , Células Hep G2 , Humanos , Separación Inmunomagnética/instrumentación , Técnicas de Diagnóstico Molecular/instrumentación , Nanopartículas/químicaRESUMEN
We describe a novel multicomponent graphene nanostructured system that is biocompatible, and has strong NIR optical absorbance and superparamagnetic properties. The fabrication of the multicomponent nanostructure system involves the covalent attachment of 3 components; Fe(3)O(4)(Fe) nanoparticles, PAMAM-G4-NH(2) (G4) dendrimer and Cy5 (Cy) on a graphene oxide (GO) surface to synthesize a biologically relevant multifunctional system. The resultant GO-G4-Fe-Cy nanosystem exhibits high dispersion in an aqueous medium, and is magnetically responsive and fluorescent. In vitro experiments provide a clear indication of successful uptake of the GO-G4-Fe-Cy nanosystem by MCF-7 breast cancer cells, and it is seen to behave as a bright and stable fluorescent marker. The study also reveals varied cellular distribution kinetics profile for the GO nanostructured system compared to free Cy. Furthermore, the newly developed GO nanostructured system is observed to be non-toxic to MDA-MB-231 cell growth, in striking contrast to free G4 dendrimer and GO-G4 conjugate. The GO-G4-Fe-Cy nanostructured system characterized by multifunctionality suggests the merits of graphene for cellular bioimaging and the delivery of bioactives.
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Neoplasias de la Mama/diagnóstico , Carbocianinas , Dendrímeros , Colorantes Fluorescentes , Grafito , Nanopartículas de Magnetita , Nylons , Carbocianinas/química , Carbocianinas/farmacocinética , Línea Celular Tumoral , Supervivencia Celular , Dendrímeros/química , Dendrímeros/farmacocinética , Femenino , Colorantes Fluorescentes/química , Colorantes Fluorescentes/farmacocinética , Grafito/química , Grafito/farmacocinética , Humanos , Nanopartículas de Magnetita/química , Microscopía Confocal , Modelos Moleculares , Nylons/química , Nylons/farmacocinética , Imagen ÓpticaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Glycosmis pentaphylla (Retz.) Correa is used in Indian traditional medicine against jaundice and other liver disorders. AIM OF THE STUDY: The study aims to determine the in vitro anticancer and apoptosis inducing activity of Glycosmis pentaphylla in hepatocellular carcinoma cell line, Hep3 B. MATERIALS AND METHODS: The cytotoxic and apoptosis inducing activity of the crude extract and active fractions were estimated on Hep3 B and RAW264.7 cell lines by MTT assay, Hoechst staining, DNA fragmentation, morphological studies, reverse transcription polymerase chain reaction and anti-poly-(ADP-ribose)-polymerase assays. The phytochemical profiling of active extract was done by TLC and HPTLC methods. RESULTS: Ethanol extract of Glycosmis pentaphylla was more effective than other extracts in reducing the proliferation of Hep3 B cells. As revealed by the results from DNA fragmentation, Hoechst staining, morphological studies, RT-PCR, PARP cleavage and gene expression studies, active extract induced apoptosis on Hep3 B cell line in concentration and time dependent manner with increase in the Bax/Bcl2 gene expression ratio. Chemo profiling data revealed the presence of flavonoid in the active fraction. CONCLUSIONS: The study showed that major active component in the ethanol extract of Glycosmis pentaphylla is a flavonoid which induces apoptosis on cancer cell line, Hep3 B, by increasing the expression ratio of Bax/Bcl2 genes in a time and dose dependent manner.