RESUMEN
NANOG is essential for mouse and human embryonic stem cell (ESC) pluripotency and selfrenewal. It is also expressed in several adult murine tissues as shown by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. However, human NANOG transcripts have been isolated from adult bone marrow (EST; GenBank accession no. BF893620). Here, we study the NANOG gene expression profile in isolated mouse renal papillary cells by Northern blot and RT-PCR. The whole RNA of mouse renal cells was obtained from fresh renal tissues, renal tissues infused by phosphate-buffered saline (PBS), and isolated renal papillary cells of mouse, respectively, as well as the renal papillary tissue from 18.5 days postcoitum (d.p.c.; fetal), 1-2-week-old (young), 1-8-month-old (adult), and 24-month-old (aging) mice. Our analysis shows that a very low expression level was detected in mouse renal tissues, and the renal papillary cells express more than other tissues as determined with Northern blot and RT-PCR. These data suggest that the kidney has its own cells expressing NANOG, and loss of NANOG expression occurs in an age-dependent manner in the kidney, either due to developmental factors or aging, particularly in renal papillary tissue.
Asunto(s)
Envejecimiento/fisiología , Proteínas de Unión al ADN/genética , Proteínas de Homeodominio/genética , Riñón/fisiología , Animales , Secuencia de Bases , Expresión Génica , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Proteína Homeótica NanogRESUMEN
OBJECTIVE: To study the expression and function of zinc ribbon gene ZNRD1 in drug-resistant cells of gastric cancer. METHODS: Two tumor cell lines were used in this study: gastric cancer SGC7901 and its drug-resistant counterpart SGC7901/VCR stepwise-selected by vincristine. The expression of ZNRD1 in SGC7901 and SGC7901/VCR was detected by northern blot and semiquantitative RT-PCR. ZNRD1 antisense nucleic acid was transfected into SGC7901/VCR cells by lipofectamine. The expression of protein in SGC7901/VCR cells and the transfectants was detected by immunochemical method. Fluorescence activated cell scan (FACS) was applied to observe the cell cycle alteration. Growth curve and drug sensitization of cells for vincristine (VCR) and adriamycin (ADM) were analyzed by MTT assay. RESULTS: The expression of ZNRD1 was higher in SGC7901/VCR than in SGC7901 cells. Immunochemical results showed that the expression level of ZNRD1 protein was lower in anti ZNRD1-SGC7901/VCR cells than in non-transfectants. The anti ZNRD1-SGC7901/VCR cells were gradually accumulated in G(1) phase, with a concomitant decrease of cell population in S phase. MTT assay showed that transfectant cell proliferation was lagged and more sensitive to VCR and ADM than non-transfectants. CONCLUSION: ZNRD1 gene displays high expression in VCR resistant gastric cancer cells. Expression of ZNRD1 protein is effectively blocked in anti ZNRD1-SGC7901/VCR cells by gene transfection. ZNRD1 antisense nucleic acid could reverse, to some degree, the MDR of human drug-resistant gastric cancer cell SGC7901/VCR.
Asunto(s)
Proteínas de Unión al ADN/fisiología , Neoplasias Gástricas/patología , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Proteínas de Unión al ADN/análisis , Proteínas de Unión al ADN/genética , Doxorrubicina/farmacología , Resistencia a Antineoplásicos , Humanos , Neoplasias Gástricas/química , Neoplasias Gástricas/tratamiento farmacológico , Vincristina/farmacologíaRESUMEN
OBJECTIVE: To study whether the porcine alpha1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte negatively expresses GT mRNA and resists to the cytotoxicity of nature antibody in human serum. METHODS: The porcine alpha1, 3 galactosyltransferase gene siRNA targeted vector (pPNTloxPGTsiRNA) were construct with pPNTloxPGT and pMXSV/U6 vector. Positive-negative selection was used to produce a heterozygous pPNTloxPGTsiRNA knockout (+/-) clone. The GT mRNA expressions were detected with northern blot. Complement-mediated NAb cytotoxicity after incubation of hepatocytes with NAbs and complement was determined using 3- (4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS, tetrazolium salt) colorimetric assay. RESULTS: The pPNTloxPGTsiRNA targeted porcine hepatocyte (+/-) negative express GT mRNA. Only 14% to 18% cytotoxicity can be detected at the highest serum concentration. The pPNTloxPGT targeted porcine hepatocyte (+/-) express GT mRNA just as the wild type porcine cells and the cytotoxicity are 77% to 83%. CONCLUSION: The porcine a1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte (+/-) negative express GT and resisted to nature antibody in human serum.