RESUMEN
Epithelial and stromal communications are essential for normal uterine functions and their dysregulation contributes to the pathogenesis of many diseases including infertility, endometriosis, and cancer. Although many studies have highlighted the advantages of culturing cells in 3D compared to the conventional 2D culture system, one of the major limitations of these systems is the lack of incorporation of cells from non-epithelial lineages. In an effort to develop a culture system incorporating both stromal and epithelial cells, 3D endometrial cancer spheroids are developed by co-culturing endometrial stromal cells with cancerous epithelial cells. The spheroids developed by this method are phenotypically comparable to in vivo endometrial cancer tissue. Proteomic analysis of the co-culture spheroids comparable to human endometrial tissue revealed 591 common proteins and canonical pathways that are closely related to endometrium biology. To determine the feasibility of using this model for drug screening, the efficacy of tamoxifen and everolimus is tested. In summary, a unique 3D model system of human endometrial cancer is developed that will serve as the foundation for the further development of 3D culture systems incorporating different cell types of the human uterus for deciphering the contributions of non-epithelial cells present in cancer microenvironment.
Asunto(s)
Comunicación Celular , Técnicas de Cocultivo , Neoplasias Endometriales/metabolismo , Neoplasias Endometriales/patología , Células Epiteliales/metabolismo , Células Epiteliales/patología , Proteómica , Comunicación Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Endometrio/diagnóstico por imagen , Endometrio/metabolismo , Endometrio/patología , Células Epiteliales/efectos de los fármacos , Everolimus/farmacología , Femenino , Hormonas Esteroides Gonadales/farmacología , Humanos , Proteoma/metabolismo , Transducción de Señal/efectos de los fármacos , Esferoides Celulares/efectos de los fármacos , Esferoides Celulares/metabolismo , Esferoides Celulares/patología , Células del Estroma/efectos de los fármacos , Células del Estroma/metabolismo , Células del Estroma/patología , Tamoxifeno/farmacologíaRESUMEN
The genome of bluetongue virus (BTV) consists of 10 segments. Of these seg-2 encoded VP2 is the major serotype determining protein, and seg-6 encoded VP5 protein enhances the protective neutralizing activity of VP2 protein inducing higher serotype specific antibody titer than the VP2 alone. Out of the twenty-six BTV serotypes found worldwide, 22 were reported from different states of India. These include serotype 21 which was recently isolated from Andhra Pradesh, and was involved in a severe outbreak of bluetongue in Indian native sheep. BTV21 (KMNO-7) and BTV16 were circulating at the same time. This co-circulation, along with the fact that the virus genome is segmented, provides an opportunity for these two isolates of different serotypes to simultaneously infect the same animal, and even the same cell or a same vector with the potential for generation of reassortant viruses. This study was carried out to provide some insights into the outbreak. We carried out full length sequencing of genome seg-2 and seg-6 of Indian isolates VJW64 (BTV16) and KMNO-7 (BTV21). Detailed phylogenetic analysis revealed that genome seg-6 of Indian isolate KMNO-7 (BTV21) clusters with isolates of BTV16 showing maximum nucleotide similarity of 97.6% with TUR/2000/02 isolate of BTV16, which is much more than it shows with any isolate of BTV21. KMNO-7 (BTV21) significantly diverged from original strain of BTV21, and is a reassortant strain having acquired seg-6 from an isolate of BTV16. This study provides some useful insights into the epidemiology of the bluetongue disease, and undermines serotyping on genome seg-6 basis.