ABSTRACT
Aim To explore the effect of shikonin on stemness maintance of glioma stem cells ( GSCs ). Methods After the U87-MG cells were cultured and isolated, the sphere cells were identified by immuno-fluorescent staining. The alteration of stemness of GSCs by shikonin treatment(2 μmol·L - 1 ) for 12 h, 24 h and 48 h was valued by morphological detection using optical microscope and sub-sphere forming assay. Mo-reover, the related markers of stem cells, such as CD133, were detected in shikonin treated GSCs by western blot assay. Protein expression of PI3K, p-PI3K, Akt and p-Akt was detected by western blot af-ter shikonin treatment alone. Furthermore, by combi-nation with insulin-like growth factor-1 ( IGF-1), we observed the alteration of stemness maintance of shiko-nin-treated GSCs. Results The presence of neural stem cell related markers CD133 and nestin proved the characteristics of GSCs. Shikonin treatment significant-ly inhibited the morphology of GSCs and the sub-sphere forming. Besides, the reduced expression of CD133 was detected in shikonin treated GSCs. Though, the expression of PI3K and Akt did not change compared with the control group, the expression of p-PI3K and p-Akt was reduced. Furthermore, the combination of IGF-1 markedly attenuated the inhibitory effect of shikonin on stemness maintance of GSCs. Conclusion The stemness maintance of GSCs can be significantly inhibited by shikonin treatment, in which PI3K/ Akt pathway is involved.
ABSTRACT
El tejido sanguíneo está compuesto en un 45% aproximadamente por células y derivados de éstas, con una vida media que oscila entre 120 días para los eritrocitos y alrededor de 3 años para ciertos tipos de linfocitos. Esta pérdida es compensada gracias a la actividad del sistema hematopoyético y a la presencia de una población de células primitivas inmaduras conocidas como Células Madre Hematopoyéticas (CMHs) encargadas del proceso de hematopoyesis, activo desde el inicio de la vida fetal y que genera cerca de 2 x 1011 eritrocitos y 1010 células blancas por día (1). Las CMHs poseen la capacidad de auto-renovarse y diferenciarse a múltiples linajes, se ubican en un nicho particular y tienen marcadores de superficie que las identifican, como por ejemplo el antígeno CD34. Recientemente se ha podido avanzar en el entendimiento de la biología básica de los procesos celulares que rigen los mecanismos de auto-renovación, diferenciación y proliferación de las CMHs, y de la participación de diferentes vías de señalización (Hedgehog, Notch y Wnt) en estos procesos, los cuales controlan el comportamiento in vivo e in vitro de las CMHs. Todo esto es de vital importancia para la implementación y generación de alternativas terapéuticas con CMHs, para diversas enfermedades entre ellas las hematológicas, como por ejemplo las leucemias.
Blood tissue is composed approximately in 45% by cells and its derivatives, with a life span of around 120 days for erythrocytes and 3 years for certain type of lymphocytes. This lost is compensated with the hematopoietic system activity and the presence of an immature primitive cell population known as Hematopoietic Stem Cells (HSCs) which perform the hematopoiesis, a process that is active from the beginning of the fetal life and produces near to 2 x 1011 eritrocytes and 1010 white blood cells per day (1). Hematopoietic Stem Cells are capable of both self-renewal and differentiation into multiple lineages, are located in a particular niche and are identified by their own cell surface markers, as the CD34 antigen. Recently it has been possible to advance in the understanding of self-renewal, differentiation and proliferation processes and in the involvement of the signaling pathways Hedgehog, Notch and Wnt. Studying the influence of these mechanisms on in vivo and in vitro behavior and the basic biology of HSCs, has given valuable tools for the generation of alternative therapies for hematologic disorders as leukemias.
Subject(s)
Humans , Hematopoiesis , Stem Cells , Hematopoietic Stem Cells , Signal Transduction , Antigens, CD34 , Hematopoietic SystemABSTRACT
Objective:To culture neural stem cell (NSC) in vitro,observe its biological property and provide basis for further study on differentiation.Methods:NSC of rat fetus were isolated and cultured.Single-cell clone culture was used to confirm the character of self-renewing.Ability of multi-directional differentiation and specific antigen, nestin,were detected by immunocytochemistry (ICC).BrdU was used to show cell stage in division.Effect of combination of EGF,bFGF on proliferation of rat NSC were evaluated.Results:The cultured NSC showed multipotential of differentiation into neurons and glial cells,as well as the ability of self-renewing.Cells expressed nestin.The population of the NSC expanded quickly and the total number of the cell increased 2.45?10~4 folds after 15 times' passage.The property of NSC was stable after long culture and preservation.Combination of EGF and bFGF stimulated cell proliferation than used singly.Conclusion:The cultured NSC express nestin, have the ability of self-renewing and multi-directional differentiation.Combination of EGF and bFGF could promote the proliferation of rat NSC.