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1.
Journal of Experimental Hematology ; (6): 525-530, 2008.
Artículo en Chino | WPRIM | ID: wpr-253283

RESUMEN

This study was aimed to explore the expression level of the unknown cgi-100 gene in human leukemia K562 cells treated with matrine, and to investigate effect of cgi-100 on proliferation of K562 cells. The expression level of cgi-100 was detected by RT-PCR in K562 cells before and after being treated with matrine; pIRES2-EGFP/cgi-100 eukaryotic expression vector was constructed by DNA recombinant technique and was introduced into K562 cells by liposome-mediated DNA transfection. The cgi-100 gene expression level, growth-curve, and cell cycle of the modified K562-cgi-100 cells were detected by RT-PCR, Trypan blue staining and FCM. Morphological changes were observed under the optical and electron microscopes. The results indicated that the expression level of cgi-100 decreased in K562 cells treated with matrine. Heterochromatin decreased, euchromatin and the proportion of S phase in K562-cgi-100 cells increased, and cell proliferation enhanced. It is concluded that the expression of cgi-100 mRNA decreased in a dose- and time-dependent manner in the K562 cells treated with matrine and over-expression of cgi-100 elevates the proliferation and the immaturity level of K562-cgi-100 cells.


Asunto(s)
Humanos , Alcaloides , Farmacología , Antineoplásicos Fitogénicos , Farmacología , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Genes Relacionados con las Neoplasias , Genética , Fisiología , Células K562 , Proteínas Proto-Oncogénicas , Genética , Metabolismo , Quinolizinas , Farmacología
2.
Chinese Journal of Hematology ; (12): 823-827, 2007.
Artículo en Chino | WPRIM | ID: wpr-262942

RESUMEN

<p><b>OBJECTIVES</b>To investigate the expression status of IER3IP1 gene during matrine induced K562 cell differentiation, and to figure out the function of IER3IP1 gene in K562 cell line.</p><p><b>METHODS</b>Trypan-blue staining was used to analyze the growth inhibitory effect of matrine on K562 cells. Semi-quantitative RT-PCR was employed to investigate the expression status of IER3IP1 gene treated with different time and dosage of matrine. The alteration of cellular morphology, cellular proliferation and ultra-microstructure were observed and the cell cycle was detected on the recombinant IER3IP1 gene eukaryotic expression vector eYFP-IER3IP1 plasmid transfected K562 cells (K562/eYFP-IER3IPl).</p><p><b>RESULTS</b>Matrine inhibited the growth of K562 cells and reduced the expression of IER3IP1 gene. The expression level of IER3IP1 gene was transiently increased to three-to-four times in a dose-dependent manner after treated with matrine for 2 - 3 hours. Then, in 6-48 hours it maintained at a low level as compared with the control group. The proliferation rate of the K562/eYFP-IER3IP1 cells significantly slowed down with more cells blocked in G0-G1 phase (P < 0.05). The number of erythroid blast cells began to increase after 24 hours of matrine treatment. At the same time, differentiated erythroid cells could be observed.</p><p><b>CONCLUSIONS</b>Matrine can inhibit the growth of K562 cells, and transiently increase the expression level of IER3IP1 gene in a dose-dependent manner. The sensitivity of K562 cells to matrine maybe increased after being transfected by the eYFP-IER3IP1 plasmid, indicating a possible involvement of the IER3IP1 gene in the early response of the cells to matrine and its possible role in the erythroid cell differentiation.</p>


Asunto(s)
Humanos , Alcaloides , Farmacología , Proteínas Portadoras , Genética , Metabolismo , Ciclo Celular , Diferenciación Celular , Genética , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Células K562 , Proteínas de la Membrana , Genética , Metabolismo , Quinolizinas , Farmacología
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