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本研究旨在观察cyclin D1、hTERT和端粒酶在正常人单个核细胞(MNC)、HL-60和HL-60A中的表达及活性,并探讨它们与白血病发生及耐药的关系。实验用正常人外周血MNC、耐药白血病细胞株HL-60和HL-60A,采用流式细胞仪检测细胞周期,Annex in V-FITC+PI染色法检测细胞凋亡,real-tim e PCR和W estern b lot检测细胞cyclin D1、hTERTmRNA及蛋白表达,TRAP-ELISA法检测细胞端粒酶活性。结果表明:MNC、HL-60、HL-60A的S期细胞比例分别为(10.21±2.11)%、(44.93±3.00)%、(51.38±1.10)%;凋亡细胞比例分别为(16.14±2.13)%、(7.53±0.92)%、(4.15±0.96)%;cyclin D1、hTERT的mRNA和蛋白水平依次增高;HL-60、HL-60A端粒酶活性较正常MNC增强(p=0.000),HL-60与HL-60A之间无明显差别(p=0.232);cyclin D1、hTERT与端粒酶活性呈正相关(p<0.01)。结论:与正常人MNC相比,HL-60、HL-60A的S期细胞增多,凋亡细胞减少,且以HL-60A更为明显,这可能与cyclin D1、hTERT、端粒酶活性上调有关。
The purpose of this study was to investigate the expression and activity of cyclin D1, hTERT and telomerase in normal human mononuclear cells (MNCs), HL-60 and HL-60A and their relationship with the development of leukemia and drug resistance. The normal human peripheral blood MNC, drug resistant leukemia cell lines HL-60 and HL-60A were used in the experiment. Cell cycle was detected by flow cytometry. Apoptosis was detected by Annexin V-FITC + PI staining. Real- The expression of cyclin D1, hTERT mRNA and protein were detected by Western blot, and telomerase activity was detected by TRAP-ELISA. The results showed that the percentage of S phase cells in MNC, HL-60 and HL-60A groups was (10.21 ± 2.11)%, (44.93 ± 3.00)% and (51.38 ± 1.10)%, respectively ), (7.53 ± 0.92)%, (4.15 ± 0.96)%, respectively. The mRNA and protein levels of cyclin D1 and hTERT increased in turn. The telomerase activity of HL-60 and HL- There was no significant difference between HL-60 and HL-60A (p = 0.232); cyclin D1 and hTERT were positively correlated with telomerase activity (p <0.01). CONCLUSIONS: Compared with normal human MNC, the number of S phase cells and the number of apoptotic cells in HL-60 and HL-60A cells are decreased and HL-60A is more obvious, which may be related to the up-regulation of cyclin D1, hTERT and telomerase activity .