RESUMO
<p><b>OBJECTIVE</b>To investigate the regulation mechanism of apoptosis induced by the antisense bcl-2 treatment.</p><p><b>METHODS</b>DNA content analysis and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) were adopted to detect apoptosis. Semi-quantitative reverse transcription-PCR was performed to detect the mRNA expression of bcl-2 c-myc survivin bax s100A(2) TNFalpha TGFbeta(1) and IL-6 in the small-cell lung cancer cell line NCI-H446 treated with antisense bcl-2 oligodeoxynucliotide.</p><p><b>RESULTS</b>bcl-2 AS-PS-ODN treatment could induce apoptosis, accompanied with 72.71% up-regulation of IL-6 and 65.90% down-regulation of TNFalpha, whereas little or no effect was seen on c-myc survivin bax s100A(2) and TGFbeta(1).</p><p><b>CONCLUSION</b>IL-6 and TNFalpha may be involved in the regulation of apoptosis induced by antisense bcl-2 treatment.</p>
Assuntos
Humanos , Apoptose , Genética , Fatores Quimiotáticos , Genética , Regulação Neoplásica da Expressão Gênica , Marcação In Situ das Extremidades Cortadas , Proteínas Inibidoras de Apoptose , Interleucina-6 , Genética , Proteínas Associadas aos Microtúbulos , Genética , Proteínas de Neoplasias , Oligonucleotídeos Antissenso , Farmacologia , Proteínas Proto-Oncogênicas , Genética , Proteínas Proto-Oncogênicas c-bcl-2 , Genética , Proteínas Proto-Oncogênicas c-myc , Genética , RNA Mensageiro , Genética , Metabolismo , Proteínas S100 , Genética , Fator de Crescimento Transformador beta , Genética , Fator de Crescimento Transformador beta1 , Células Tumorais Cultivadas , Fator de Necrose Tumoral alfa , Genética , Proteína X Associada a bcl-2RESUMO
AIM: To investigate the relationship between the expressions of DNA topoisomerase (Topo), glutathione S-transferases (GSTs) and chemotherapy response, prognosis in acute leukemia (AL). METHODS: Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of TopoⅠ, Ⅱ?, Ⅱ? and GST?, ? from patients with AL. RESULTS: The results showed that the relative mRNA expression level of TopoⅡ?, TopoⅡ? and GST ? in AL group were significantly higher than that in normal subjects. GST?, however, was exactly reverse ( P
RESUMO
AIM:To investigate the effects of baicalin on proliferation inhibition and apoptosis induction in human Burkitt lymphoma cell line CA46 and to explore its underlying mechanisms.METHODS:CA46 cells were exposed to baicalin at different dosages and its proliferation inhibition was detected by MTT assay.The ability of baicalin to induce CA46 cell apoptosis was examined by Annexin V-FITC/PI double staining analysis,TUNEL labeling method and DNA fragmentation.The mRNA expressions of c-myc and bcl-2 were detected by RT-PCR,and the protein expressions of c-Myc,Bcl-2,caspase-3 precursor(procaspase-3) and poly ADP-ribose polymerase(PARP) were detected by Western blotting.RESULTS:Baicalin remarkably inhibited the CA46 cell proliferation,with an IC50 value of 10 ?mol/L.Apoptosis was remarkably induced by baicalin in a dose-dependent manner,and its earlier and later stages were detected by annexin V-FITC/PI double staining analysis,TUNEL labeling method and DNA fragmentation,respectively.Furthermore,RT-PCR showed that the mRNA expressions of c-myc and bcl-2 in treated CA46 cells decreased in a time-dependent manner.Western blotting showed that the protein expressions of c-Myc,Bcl-2,procaspase-3 and PARP(116 kD) in baicalin treated CA46 cells were down-regulated in a time-dependent manner,while the expression of PARP(85 kD) was up-regulated.CONCLUSION:Baicalin efficiently induces proliferation inhibition and apoptosis in CA46 cells,which may be related with the down-regulation of c-Myc and Bcl-2 expressions,as well as the up-regulation of caspase-3 activity.
RESUMO
AIM: To observe the effect of antisense bcl-2 oligodeoxynucleotides(AS-PS-ODN) on bcl-2 mRNA and protein expression, cell proliferation,viability and apoptosis in a small-cell lung cancer cell line NCI-H446. METHODS: Semi-quantitative RT-PCR was performed to detect the bcl-2 mRNA expression, the Bcl-2 protein was determined by immunocytochemistry and flow cytometry analysis, and the effect of bcl-2 AS-PS-ODN on cell proliferation, viability and apoptosis were investigated by colony assay , cell count, DNA content analysis and TUNEL. RESULTS: ① 1 ?mol/L bcl-2 AS-PS-ODN significantly down-regulated the expression of bcl-2 mRNA and protein. The inhibition rate of mRNA and protein were 69.5% and 62.7%, respectively. ② bcl-2 AS-PS-ODN decreased cell proliferation and viability , induced cell apoptosis.The apoptosis rate was 22.3%-32.7% in cells treated with 1?mol/L bcl-2 AS-PS-ODN. CONCLUSION: bcl-2 AS-PS-ODN down-regulated expression of bcl-2 mRNA and protein, inhibited cell proliferation and induced apoptosis in a small cell lung cancer cell line, NCI-H446.
