摘要
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。
To observe the expression of cyclin D1, hTERT, and telomerase activity in MNC, HL-60, HL-60A and to explore their effects on leukemogenesis and drug-resistance, normal human peripheral blood mononuclear cells, HL-60 cells sensitive to adriamycin and HL-60A cells resistant to adriamycin were investigated. The cell cycle was analyzed by flow cytometry, and the apoptosis was analyzed by Armexin V-FITC＋ PI staining. Expressions of cyclin D1 and hTERT were determined by real-time PCR and Western blot. Telomerase activity was detected by TRAP-ELISA. The results indicated that the percentage of MNC, HL-60 and HL-60A in S phase was （10.21 ±2.11）% ,（44.93 ± 3.00）%, and （51.38 ± 1.10） % respectively; the percentage of apoptosis cells was （ 16.14 ± 2.13 ） %, （7.53 ± 0.92） %, （4.15 ± 0.96） % respectively; the expression of mRNA and protein for cyclin D1 and hTERT increased; the telomerase activities of HL-60 and HL-60A were higher（p = 0.000） ,whereas the difference between HL-60 and HL-60A was no statistically significant （p = 0.232）; positive correlation between cyclin D1, hTERT and telomerae activity had been found （p 〈 0.01 ）. It is concluded that the cells of S phase increased while the apoptotic cells decreased in HL-60 and HL-60A, especially in HL-60A,which may be due to the up-regulation of cyclin D1, hTERT and telomerase activity.
出处
《中国实验血液学杂志》
CAS
CSCD
2011年第4期911-915,共5页
Journal of Experimental Hematology
基金
广东省自然科学基金(编号9151008901000105)
广东省科技计划项目(编号2008B060600032)
