RESUMO
The study was aimed to investigate the possible mechanism of vitamin K(2) (VK(2)) on myelodysplastic syndrome (MDS) cell line MUTZ-1 in vitro. The flow cytometry was used to analyze apoptosis rate and the change of cell cycle. The expression of apoptosis-related genes bcl-2, survivin and bax were detected by reverse transcription-polymerase chain reaction (RT-PCR). The activity of caspase-3 was detected by chemiluminescence assay. The results indicated that the apoptosis peak on FCM and positive Annexin-V FITC on cell membrane showed that VK(2) induced apoptosis of MUTZ-1 cells in a dose-and-time-dependent manner, S and G(2) cell decrement, G(0)/G(1) cell arrest, VK(2) significantly down-regulated the expression of bcl-2 and survivin, but had no effect on the expression of bax, the activity of caspase-3 was significantly increased. It is concluded that VK(2) induces apoptosis of MUTZ-1 cells through activating caspase-3 pathways and the apoptosis-related genes bcl-2 and survivin may play important roles in the process of apoptosis induction.
Assuntos
Humanos , Antineoplásicos , Farmacologia , Apoptose , Caspase 3 , Metabolismo , Linhagem Celular Tumoral , Proteínas Inibidoras de Apoptose , Proteínas Associadas aos Microtúbulos , Genética , Síndromes Mielodisplásicas , Tratamento Farmacológico , Patologia , Proteínas de Neoplasias , Genética , Proteínas Proto-Oncogênicas c-bcl-2 , Genética , RNA Mensageiro , Genética , Vitamina K 2 , Farmacologia , Proteína X Associada a bcl-2 , GenéticaRESUMO
To study the effects and possible mechanism of Vitamin K(2) (VK(2)) in the treatment of MDS-JSN04 cells, the changes of morphologic features of MDS-JSN04 cells were investigated by cytomorphology, the apoptosis of MDS-JSN04 cells was observed by transmission electron microscope; cellular proliferation was determined by the MTT assay; cell apoptosis, cell cycle shift and expression of myeloid-specific differentiation antigen (CD11b, CD13) were analyzed by flow cytometry (FCM). The expression of apoptosis-related genes bcl-2, survivin and bax were detected by retrotranscriptase polymerase chain reaction (RT-PCR); the activity of caspase-3 was determined by chemiluminescence assay. The results showed that the typical apoptotic morphological features appeared in cells treated with VK(2) for 72 hours; VK(2) induced apoptosis of MDS-JSN04 cells and in a dose-and-time-dependent manner, G(0)/G(1) cell arrest and significantly down-regulated the expression of bcl-2 and survivin, but had no effect on the expression of bax; the activity of caspase-3 significantly increased. It is concluded that VK(2) induces apoptosis of MDS-JSN04 cells through activating caspase-3 pathways and the apoptosis-related genes bcl-2, survivin may play an important role in this process.
Assuntos
Humanos , Apoptose , Antígeno CD11b , Antígenos CD13 , Caspase 3 , Metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Citometria de Fluxo , Regulação Neoplásica da Expressão Gênica , Proteínas Inibidoras de Apoptose , Medições Luminescentes , Métodos , Microscopia Eletrônica de Transmissão , Proteínas Associadas aos Microtúbulos , Genética , Síndromes Mielodisplásicas , Genética , Metabolismo , Patologia , Proteínas de Neoplasias , Genética , Proteínas Proto-Oncogênicas c-bcl-2 , Genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Métodos , Vitamina K 2 , Farmacologia , Proteína X Associada a bcl-2 , GenéticaRESUMO
This study was aimed to establish a cytokine-independent human myelodysplastic cells line from bone marrow of a patient with MDS-CMML. This cell line was incubated in mixed culture of RPMI 1640 and DMEM with 15% bovine serum, but without cytokines; its biological characteristics were identified by morphology, surface marker profiles, cell proliferation, differentiation and apoptosis. The results showed that the established cell line could not depend on cytokines for long-term survival and growth, and could differentiate into colony-forming unit-macrophage, colony-forming unit-megakaryocyte. In conclusion, a cytokine-independent human myelodysplastic syndrome cell line, named MDS-JSN04 (MDS Nanjing Jiansu 04), was established. Its partial biological characteristics were identified and clarified.