Yang Jian-Hui, Zhang Yi-Chu, Qian Hui-Qing
Department of Oncological Surgery, Shidong Hospital, Ningguo Road 236, Shanghai 20090, China.
World J Gastroenterol. 2004 Apr 15;10(8):1121-4. doi: 10.3748/wjg.v10.i8.1121.
To investigate the effect of transfected survivin antisense oligonucleotide (ASODN) on proliferation and apoptosis of gastric cancer cells.
The authors designed ASODNs targeting different regions of survivin mRNA, including surviving ASODN1, ASODN2 and ASODN3. ASODNs were transfected into gastric cancer cell line SGC 7901, cell growth was detected by MTT assay. Cells exposed to the potent oligonucleotide were also examined for apoptosis induction by FCM and fluorescence microscopy. Semiquantitive RT-PCR and Western blot examinations were carried for expression of survivin mRNA and protein.
ASODN3 caused a statistically significant reduction of cell viability to 60.6% (+/-2.9%) (P<0.01), while ASODN1 and ASODN2 had no such changes (P>0.05). The cell growth was also significantly inhibited by ASODN3, compared with reversal and scrambled sequence. A significant loss of survivin mRNA was presented in ASODN3 treated cells and this was not seen in treatment with sense ODN or scramble ODN. Protein level was significantly decreased 48 h after survivin ASODN trasfected by approximately 2-fold decrease compared with untreated controls. However, ASODN3 did not induce significant apoptosis response until 48 h after transfection (P>0.05).
ASODN3, which targets translation initiation part, can be identified as a most potent antisense compound. Srvivin ASODN3 may provide a novel approach to therapy of gastric cancer.
探讨转染生存素反义寡核苷酸（ASODN）对胃癌细胞增殖和凋亡的影响。
作者设计了针对生存素mRNA不同区域的ASODN，包括生存素ASODN1、ASODN2和ASODN3。将ASODN转染至胃癌细胞系SGC 7901，采用MTT法检测细胞生长情况。通过流式细胞术（FCM）和荧光显微镜检查暴露于有效寡核苷酸的细胞的凋亡诱导情况。进行半定量逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹法检测生存素mRNA和蛋白质的表达。
ASODN3使细胞活力在统计学上显著降低至60.6%（±2.9%）（P<0.01），而ASODN1和ASODN2无此变化（P>0.05）。与正义寡核苷酸和乱序序列相比，ASODN3也显著抑制细胞生长。在ASODN3处理的细胞中出现生存素mRNA的显著缺失，而在正义寡核苷酸或乱序寡核苷酸处理中未见此现象。在转染生存素ASODN 48小时后，蛋白质水平与未处理对照相比显著降低约2倍。然而，ASODN3直到转染后48小时才诱导出显著的凋亡反应（P>0.05）。
靶向翻译起始部分的ASODN3可被确定为最有效的反义化合物。生存素ASODN3可能为胃癌治疗提供一种新方法。
