CAJ | 학술논문

目的观察全硫代反义寡聚脱氧核苷酸(PS-ASODN)对胃肠道间质瘤细胞株GIST867增殖、凋亡及端粒酶活性的影响.方法将1μmol/L伊马替尼和1.25、2.50、5.00、10.00、20.00 μmol/L PS-ASODN作用于GIST867；采用噻唑蓝(MTT)比色法、酶联免疫吸附试验(ELISA)和流式细胞术检测增殖抑制、端粒酶活性和细胞凋亡,应用逆转录-聚合酶链反应(RT-PCR)检测B淋巴细胞/白血病-2(bcl-2) mRNA表达.结果 MTT显示PS-ASODN浓度为5.00μmol/L时,抑制作用明显较对照组强；PS-ASODN对端粒酶活性有明显抑制作用并具时间依赖性；流式细胞检测表明PS-ASODN组的细胞凋亡率为(16.16±1.35)％,显著高于伊马替尼组及对照组；RT-PCR检测表明PS-ASODN可显著下调bcl-2 mRNA的表达.结论 PS-ASODN可能通过抑制端粒酶活性并下调bcl-2基因表达从而抑制GIST867细胞增殖及诱导细胞凋亡.
목적 관찰전류대반의과취탈양핵감산(PS-ASODN)대위장도간질류세포주GIST867증식、조망급단립매활성적영향.방법 장1μmol/L이마체니화1.25、2.50、5.00、10.00、20.00 μmol/L PS-ASODN작용우GIST867；채용새서람(MTT)비색법、매련면역흡부시험(ELISA)화류식세포술검측증식억제、단립매활성화세포조망,응용역전록-취합매련반응(RT-PCR)검측B림파세포/백혈병-2(bcl-2) mRNA표체.결과 MTT현시PS-ASODN농도위5.00μmol/L시,억제작용명현교대조조강；PS-ASODN대단립매활성유명현억제작용병구시간의뢰성；류식세포검측표명PS-ASODN조적세포조망솔위(16.16±1.35)％,현저고우이마체니조급대조조；RT-PCR검측표명PS-ASODN가현저하조bcl-2 mRNA적표체.결론 PS-ASODN가능통과억제단립매활성병하조bcl-2기인표체종이억제GIST867세포증식급유도세포조망.
Objective To explore effects of the telomerase RNA-targeting antisense oligonucleotides PS-ASODN on proliferation,apoptosis and telomerase activity of the gastrointestinal stromal tumor cell line GIST867 cultured in vitro.Methods GIST867 cells in exponential phase were divided into following groups:control group,1 μmol/L imatinib group,and 1.25,2.50,5.00,10.00 and 20.00 μmol/L PSASODN groups.The anti-proliferation effect on GIST867 cells was determined by methyl thiazol tetrazolium (MTT) assay. The activity of telomerase was tested by using enzyme linked immunosorbent assay (ELISA).Apoptosis was examined by using flow cytometry.Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of B lymphocytes/leukemia-2 (bcl-2) mRNA.Results MTT assay showed PS-ASODN could apparently inhibit the growth of GIST867 cells in a dose-dependent manner.The group of 5.00 μmol/L concentration showed more effectiveness than control group. PSASODN could apparently inhibit the telomerase activity in a time-dependent manner.Flow cytometry revealed that the apoptosis rate of the cells treated by PS-ASODN was ( 16.16 ± 1.35 ) ％,significantly higher than that in the control group and the imatinib group.Treatment with PS-ASODN significantly reduced the bcl-2 mRNA expression in the GIST867 cells.Conclusion PS-ASODN can repress the cell proliferation and induce the apoptosis of the GIST867 cells,which may be mediated by inhibition of the telomerase activity and down-regulation of the expression of bcl-2.