RESUMEN
To investigate the effect of cyclin G1 antisense oligodeoxynucleotide (ASON) with liposomal transfection on mediating proliferation of HL-60 cell, the cyclin G1 ASON with liposomal transfection was used in vitro in co-culture with HL-60 cell, the protein and mRNA expression levels of cyclin G1 were measured by immunocytochemistry assay and RT-PCR. The cell apoptosis was detected by electron microscopy, in situ cell apoptosis detection kit (POD), DNA gel electrophoresis and flow cytometry (FCM). The results showed that in the cyclin G1 ASON group the protein and mRNA expression of cyclin G1 were significantly inhibited as compared with sense oligodeoxynucleotide (SON) group and blank group. When the ASON concentration increased, the proliferation ratio of HL-60 cell and CFU of HL-60 were also significantly inhibited. There was apoptosis of HL-60 cell. In conclusion, cyclin G1 ASON can specifically inhibit its protein and mRNA expression levels as well as the HL-60 cell proliferations and can accelerate the apoptosis of leukemia cells with concentration-dependent effect of ASON.
Asunto(s)
Ciclinas/antagonistas & inhibidores , Células HL-60/efectos de los fármacos , Oligonucleótidos Antisentido/farmacología , Apoptosis/efectos de los fármacos , División Celular/efectos de los fármacos , Ciclina G , Ciclina G1 , Ciclinas/genética , Citometría de Flujo , Células HL-60/citología , Humanos , Liposomas , Microscopía Electrónica , TransfecciónRESUMEN
OBJECTIVE: To explore the effect of liposomal transfection of cyclin A antisense oligodeoxynucleotide (ASON) on HL-60 cell proliferation and apoptosis. METHODS: By liposomal transfection, cyclin A ASON was co-cultured with HL-60 cells, the cell growth curve was determined by MTT assay and cell apoptosis electron-microscopy in situ cell apoptosis detection kit (POD), the protein and mRNA of cyclin A and bcl-2 were measured by FACS and RT-PCR, the role of cyclin A ASON in the development of leukemia was tested by the tumor formation in nude mice. RESULTS: (1) In the cyclin A ASON liposomal transfection group (group A), the proliferation of HL-60 cell was significantly inhibited as compared to those in cyclin A ASON group (group B) (68.9% vs 24.8%) (P < 0.01). (2) The expressions of cyclin A and bcl-2 of group A were significantly lower than those in the control group (1.1% vs 38.8%, P < 0.01; 21.9% vs 65.0%, P < 0.01, respectively), and the DNA ladder and apoptosis body was displayed. (3) In group A, the rate of tumor formation in nude mice was lower, the time for tumor formation was longer and the volume of tumor was smaller than those in control group. CONCLUSION: Liposomal transfection of cyclin A ASON can inhibit in vitro proliferation of leukemia cells and induce in vivo apoptosis of the tumor cell, which might provide a new target for gene therapy.
Asunto(s)
Apoptosis/efectos de los fármacos , Ciclina A/fisiología , Oligonucleótidos Antisentido/farmacología , Animales , División Celular/efectos de los fármacos , Ciclina A/genética , Terapia Genética , Células HL-60 , Humanos , Leucemia/terapia , Liposomas , Ratones , Ratones Endogámicos BALB C , TransfecciónRESUMEN
OBJECTIVE: To investigate the expression of cyclins related gene in HL60 cells before and after arsenic trioxide treatment by gene chip. METHODS: Total mRNAs were extracted from untreated or arsenic trioxide treated HL60 cells. Then two cDNA probes were made from these two mRNAs which were labeled by fluoro link Cy3-ducpp deoxyuridine triphosphate (Cy3-dUTP) or Cy5-dUTP fluorescence dyes respectively, hybridized with gene chip and scanned for fluorescent intensity. Different expression genes were then screened out. Cell apoptosis was detected by electron microscopy, in site cell apoptosis detection kit, DNA agarose gel electrophoresis and flow cytometry (FCM). RESULTS: Among the 82 genes which expressed differently after treatment with arsenic trioxide, 34 genes were up-regulated, while 48 genes down-regulated. It was detected that 15 micro mol/L As(2)O(3) can definitively induce HL60 cells to go apoptosis by FCM. Rate of apoptotic HL60 cells in control group is 1.7%, and As(2)O(3) group is 26.1%. CONCLUSION: Cyclin B1, proliferating cell nuclear antigen (PCNA), insulin like growth factor binding protein (IGFBP) et al may play an important role in HL60 cell apoptosis induced by arsenic trioxide.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Arsenicales/farmacología , Ciclinas/genética , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Óxidos/farmacología , Trióxido de Arsénico , Ciclo Celular/efectos de los fármacos , Regulación hacia Abajo , Perfilación de la Expresión Génica , Células HL-60 , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Regulación hacia ArribaRESUMEN
OBJECTIVE: To study the inhibition effect of cyclin G(1) antisense oligodeoxynucleotides (ASON) on the growth of HL-60 cells in nude mice. METHODS: (1) Nude mice were divided into control group, sense oligodeoxynucleotides (SON) group and ASON group. After (60)Co radiation, with HL-60 cells SON group and ASON group were subcutaneously innoculated; (2) The weight and volume of tumors were continually measured; (3) The morphology of tumor cells was observed by microscope; (4) The protein and mRNA expression levels of cyclin G(1) were determined by flow cytometry (FCM) and reverse transcription polymerase chain reaction (RT-PCR); (5) The cell apoptosis was detected by electron microscopy and FCM. RESULTS: (1) The inhibition rate of tumor in ASON group was 69.4%. In ASON group, the wight and volume of tumor were significantly lower than those in SON group and control group. (2) The HL-60 cells in ASON group showed morphologically smaller nuclei, less mitosis, less heteromorphosis and apoptosis. CONCLUSION: The cyclin G(1) ASON can inhibit the growth of HL-60 cells in nude mice and induce apoptosis.
