He D, Zhang H
Institute of Hematology, Jinan University Medical College, Guangzhou 510632, China.
Zhonghua Nei Ke Za Zhi. 2001 Oct;40(10):654-6.
To explore the effect of human telomerase reverse transcriptase(hTERT) gene antisense oligodeoxynucleotide (ASODN) on cisplatin-induced apoptosis in cultured primary acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) cells.
Cell surviving fraction was determined using the trypan blue dye exclusion assay. Apoptosis was detected by DNA gel electrophoresis and flow cytomertric cell cycle analysis.
Treatment with cisplatin after 24 hours of exposure of the cells to ASODN significantly reduced the number of viable AML and CML cells. However, there was no difference on AML and CML cells survival between those treated with sense oligodeoxynucleotide (SODN)/cisplatin combination and cisplatin alone. Agarose gel electrophoresis of genomic DNA from the blasts of AML and CML treated with ASODN and cisplatin combination for 72 hours showed typical DNA "ladder", but DNA from AML or CML cells treated with SODN plus cisplatin or cisplatin alone did not. The percentage of apoptotic AML and CML cells treated with cisplatin for 48 or 72 hours after 24 hours of exposure to ASODN (42.68%, 35.72%) was significantly different with that of apoptotic cells treated with SODN plus cisplatin (29.02%, 23.84%) or cisplatin alone (27.53%, 21.02%).
hTERT ASODN could enhance the cisplatin-induced apoptosis of AML and CML cells.
探讨人端粒酶逆转录酶(hTERT)基因反义寡脱氧核苷酸(ASODN)对顺铂诱导培养的原发性急性髓系白血病(AML)和慢性髓系白血病(CML)细胞凋亡的影响。
采用台盼蓝拒染法测定细胞存活分数。通过DNA凝胶电泳和流式细胞术细胞周期分析检测细胞凋亡。
细胞暴露于ASODN 24小时后再用顺铂处理,可显著减少存活的AML和CML细胞数量。然而,用正义寡脱氧核苷酸(SODN)/顺铂联合处理组与单用顺铂处理组的AML和CML细胞存活率无差异。ASODN与顺铂联合处理AML和CML细胞72小时后,基因组DNA的琼脂糖凝胶电泳显示典型的DNA“梯状”条带,但SODN加顺铂或单用顺铂处理的AML或CML细胞的DNA未出现此现象。细胞暴露于ASODN 24小时后,用顺铂处理48或72小时的凋亡AML和CML细胞百分比(42.68%,35.72%)与用SODN加顺铂(29.02%,23.84%)或单用顺铂(27.53%,21.02%)处理的凋亡细胞百分比有显著差异。
hTERT ASODN可增强顺铂诱导的AML和CML细胞凋亡。