摘要
目的 探讨人端粒酶催化亚基 (hEST2基因 )反义寡核苷酸 (AODN)对卵巢癌细胞系SKOV3、COC1细胞端粒酶活性及细胞生长的影响。方法 分别采用逆转录聚合酶链反应技术、端粒重复扩增法、四甲基偶氮唑蓝法、绘制细胞生长曲线的方法检测hEST2基因AODN转染前后SKOV3、COC1细胞hEST2mRNA表达、端粒酶活性的变化 ,以及对细胞增殖能力及细胞生长的影响。结果hEST2基因AODN能够下调SKOV3、COC1细胞hEST2mRNA的表达 ,降低端粒酶活性 ,抑制细胞的生长 ,作用具有明显的时效性。以浓度为 30 μmol/L的AODN治疗 4 8h时作用最显著 ,SKOV3、COC1细胞hEST2mRNA的表达分别下降 5 4 6 %、4 4 6 % ,对两株细胞的端粒酶活性抑制率分别达到 4 7 9%、4 2 7% ,与相同浓度的随机寡核苷酸 (RODN)、正义寡核苷酸 (SODN)相比 ,差异有显著性 (P <0 0 5 )。结论 hEST2基因的反义治疗能够抑制卵巢癌细胞的增殖能力 。
Objective To study the effects of antisense oligodeoxynucleotides(ODN) of hEST2 (AODN) on telomerase activity and proliferation in ovarian cancer cell lines SKOV3 and COC1 Methods Antisense and sense human telomerse catalytic sub unit (hEST2) phosphorothioate (SODN)and random ODN were designed, synthesized and transfected into SKOV3 and COC1 cells by lipofectamine The expression of hEST2 mRNA and telomerase activity in SKOV3 and COC1 were tested by reverse transcription polymerase chain reaction and telomeric repeat amplification protocol before and after transfection The proliferation and growth in SKOV3 and COC1 were also investigated by methyl thiazolyl tetrazolium and growth curve before and after transfection Results AODN could down regulate the expression of hEST2 mRNA, inhibit telomerase activity and proliferation of ovarian cell lines The efficiency depends on dose and period of administration At 48 h, 30 μmol/L AODN had the highest activity The expression of hEST2 mRNA were declined 54 6% and 44 6% in SKOV3 and COC1 respectively And also the inhibition of telomerase activity were 47 9% and 42 7% respectively in the two cell lines Conclusions AODN of hEST2 clearly inhibited the proliferation of ovarian cancer cell lines hEST2 may thus be a new target of gene therapy in ovarian carcinoma
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2002年第4期198-201,共4页
Chinese Journal of Obstetrics and Gynecology
