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Objective To investigate the expression profile of Survivin gene in hematological tumour cell lines. Method The transcriptional levels of Survivin gene were detected by RT-PCR. Results Survivin gene was expressed in 11 hematological tumout cell lines(K562,HL-60,U937,HEL,6T-CEM,NB4,Jurkat,Raji,MUTZ-1,Dami and K562/ADR cell lines), in which the transcriptional levels were significantly higher than those in disease controls. In addition, there existed differential expression levels of Survivin gene in hematological... Objective To investigate the expression profile of Survivin gene in hematological tumour cell lines. Method The transcriptional levels of Survivin gene were detected by RT-PCR. Results Survivin gene was expressed in 11 hematological tumout cell lines(K562,HL-60,U937,HEL,6T-CEM,NB4,Jurkat,Raji,MUTZ-1,Dami and K562/ADR cell lines), in which the transcriptional levels were significantly higher than those in disease controls. In addition, there existed differential expression levels of Survivin gene in hematological tumour cell lines(NB4>MUTZ-1>K562>Raji>HL-60>6T-CEM>U937>HEL>Jurkat>Dami).The transcriptional levels of Survivin gene in K562/ADR cell line increased 1.6-fold compared with the parent K562 cell line. The Survivin expression was not detected in peripheral blood mononuclear cells. All-trans retinoic acid significantly decreased Survivin mRNA expression levels. Conclusion The Survivin gene might be a potential therapeutic target due to the role in anti-apoptosis and drug resistance. 目的 探讨 Survivin基因在血液肿瘤细胞株细胞的表达及其可能的机制。方法 应用 RT- PCR半定量法检测 Survivin基因 m RNA表达。结果 血液肿瘤细胞株和 K5 6 2 / ADM细胞株细胞均有 Survivin m RNA表达 ,且均高于疾病对照组 (P<0 .0 0 1)。不同的血液肿瘤细胞株 Survivin m RNA表达有明显的差异性 ,Dami与K5 6 2、HL - 6 0、Jurkat、MU TZ- 1、Raji、6 T- CEM;NB4与 U 937、HEL、6 T- CEM、Jurkat、Dami;HEL与 MU TZ- 1之间Survivin m RNA的表达差异有显著性 (P值分别 <0 .0 0 1~ 0 .0 5 )。 K5 6 2 / ADM细胞株与 K5 6 2细胞株细胞相比 ,Survivin m RNA表达上调了 1.6倍。外周血单个核细胞 Survivin m RNA表达阴性。经 ATRA处理后 ,NB4细胞Survivin m RNA表达随着时间的延长而呈逐渐下降趋势 ,在 72小时时几乎检测不到 Survivin... 目的 探讨 Survivin基因在血液肿瘤细胞株细胞的表达及其可能的机制。方法 应用 RT- PCR半定量法检测 Survivin基因 m RNA表达。结果 血液肿瘤细胞株和 K5 6 2 / ADM细胞株细胞均有 Survivin m RNA表达 ,且均高于疾病对照组 (P<0 .0 0 1)。不同的血液肿瘤细胞株 Survivin m RNA表达有明显的差异性 ,Dami与K5 6 2、HL - 6 0、Jurkat、MU TZ- 1、Raji、6 T- CEM;NB4与 U 937、HEL、6 T- CEM、Jurkat、Dami;HEL与 MU TZ- 1之间Survivin m RNA的表达差异有显著性 (P值分别 <0 .0 0 1~ 0 .0 5 )。 K5 6 2 / ADM细胞株与 K5 6 2细胞株细胞相比 ,Survivin m RNA表达上调了 1.6倍。外周血单个核细胞 Survivin m RNA表达阴性。经 ATRA处理后 ,NB4细胞Survivin m RNA表达随着时间的延长而呈逐渐下降趋势 ,在 72小时时几乎检测不到 Survivin m RNA的表达。结论 Survivin基因是肿瘤细胞抗凋亡和耐药的重要机制之一 ,Survivin基因有望成为肿瘤治疗新靶点 Aim To explore whether the oligonucleotide upkake in hematological tumor cells is related to cellular species and proliferation. Methods Intracellular mean fluorescence intensity was measured by flow cytometry. Results After treatment with FITC-labeled G3139 at the concentration of 0 60 μmol·L -1 for 4 h, the G3139 uptake into peripheral blood mononuclear cell and bone marrow mononuclear cell in hematological tumor patients was significantly higher than that in normal control. There was different uptake... Aim To explore whether the oligonucleotide upkake in hematological tumor cells is related to cellular species and proliferation. Methods Intracellular mean fluorescence intensity was measured by flow cytometry. Results After treatment with FITC-labeled G3139 at the concentration of 0 60 μmol·L -1 for 4 h, the G3139 uptake into peripheral blood mononuclear cell and bone marrow mononuclear cell in hematological tumor patients was significantly higher than that in normal control. There was different uptake of G3139 among the malligant hematological tumor cell strains, and the uptake in cells derived from monocyte, B lymphocyte and myeloid cell was much higher than that in cells derived from T lymphocyte. After treatment with all-trans retinoic acid (ATRA), HL60 cell proliferation was markedly inhibited and the uptake of G3139 decreased significantly. Conclusion Hematological tumor cells were capable of taking up oligonucleotide, and the oligonucleotide uptake in hematological tumor cells is related to its cellular species and its activation. 目的 探讨血液肿瘤细胞摄取寡核苷酸与其种类和活性的关系。方法 流式细胞仪测定细胞内的平均荧光强度。结果 经过 0 60 μmol·L-1 荧光素FITC标记的G31 39作用 4h后 ,血液系统肿瘤患者外周血及骨髓单个核细胞对G31 39的摄取能力明显高于正常人 ;不同来源的血液肿瘤细胞株摄取G31 39的能力不同 ,单核细胞、B淋巴细胞和髓系粒细胞来源的白血病细胞的摄取能力明显高于T淋巴细胞来源的白血病细胞 ;经过全反式维甲酸作用的HL60细胞的增殖活性明显受到抑制 ,同时细胞摄取G31 39的能力明显下降。结论 血液肿瘤细胞具有摄取寡核苷酸的能力 ,这种摄取能力与其细胞种类和细胞增殖活性有关 Aim: To detect telomerase activity in malignant hematological cells by polymerase chain reaction enzyme-linked immunoassay (PCR-ELISA) and polymerase chain reaction-polyacrylamide gel electrophoresis (PCR-PAGE), and to investigate their clinical signifcance. Methods: The telomerase PCR-ELISA is an assay in which telomeric repeat amplification products were detected by ELISA. The PCR-PAGE assay uses polyacrylamide gel electrophoresis to determine telomeric repeat amplification products. Results: A linear relationship... Aim: To detect telomerase activity in malignant hematological cells by polymerase chain reaction enzyme-linked immunoassay (PCR-ELISA) and polymerase chain reaction-polyacrylamide gel electrophoresis (PCR-PAGE), and to investigate their clinical signifcance. Methods: The telomerase PCR-ELISA is an assay in which telomeric repeat amplification products were detected by ELISA. The PCR-PAGE assay uses polyacrylamide gel electrophoresis to determine telomeric repeat amplification products. Results: A linear relationship between the absorbance and the number of PCR cycles was obtained within 25~35 cycles. A typical plateau effect reached after 35 PCR cycles. HL-60, K562, Raji cells were found to express high levels of telomerase activity by PCR-ELISA assay in which the absorbance were 2362, 2336, 2145, respectively. The telomeric repeat amplification products by PAGE-sliver staining showed a typical ladder with the telomerase-specific 6-nucleotide increments. WConclusion:ELISA and PAGE-sliver staining in the meantime can show telomeric repeat amplification products. Both of them could be suitable for clinical application, but PCR-ELISA assay is more simple and sensitive. 目的:通过端粒酶聚合酶链反应-酶联免疫测定(PCR-ELISA)和聚合酶链反应-聚丙烯酰胺凝胶电泳(PCR-PAGE)方法检测血液肿瘤细胞株HL-60、K562、Raji的端粒酶活性,探讨其临床应用价值。方法:PCR-ELISA方法是将端粒重复序列PCR变性产物与地高辛标记且对端粒重复片段特异的探针杂交,杂交产物通过ELISA进行测定。PCR-PAGE法是将端粒重复序列PCR产物直接进行聚丙烯酰胺凝胶电泳,通过硝酸银染色测定端粒酶活性。结果:在端粒酶PCR-ELISA方法检测中,PCR在25~35次循环之间,吸光度A值与循环次数呈线性关系,35~45次循环已达到平台期。PCR-ELISA法检测显示HL-60、K562、Raji细胞均表达高水平的端粒酶活性,吸光度A均值分别为2 362、2 336、2 145。对HL-60、K562、Raji细胞的端粒重复序列扩增的PCR产物直接进行PAGE电泳、银染均呈现3条以上间隔6bp的特征性阶梯状条带。结论:对端粒重复序列PCR产物可同时进行ELISA检测和PAGE-银染,两种方法均可用于恶性肿瘤的临床诊断,ELISA法更为灵敏而简单,且可相对定量。
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