摘要
[目的]观察survivin反义寡核苷酸(ASODN)对HT29细胞增殖和凋亡的影响。[方法]针对sur- vivin基因序列设计合成survivin反义寡核苷酸并通过脂质体将其转染至HT29细胞中。噻唑蓝(MTT)法观察survivin ASODN对HT29细胞的细胞毒作用,测定IC_(50);Hoechst33342/PI双荧光染色后,观察转染survivin A- SODN后HT29细胞核变化;流式细胞仪检测细胞周期。[结果]200、400、600、800、1 000及1 200 nmol/L survivin ASODN分别处理HT29细胞44 h后,Ic_(50)值为1 000 nmol/L。200、400、600、800和1 200 nmol/L组抑制率分别为(12.38±7.96)%、(22.30±4.49)%、(26.51 4±3.08)%、(38.90±3.66)%和(63.97±4.50)%。1 000 nmol/L survivin ASODN处理HT29细胞36 h后,经Hoechest33342/PI双荧光染色可观察到染色质高度浓缩、边缘化,凝集成明亮的团块。流式细胞仪检测1 000 nmol/L survivin ASODN处理HT29细胞36 h后,G_2期细胞显著增多。[结论]Survivin ASODN可显著抑制HT29细胞增殖,诱导其凋亡。Survivin靶向治疗可能成为结肠癌综合治疗的一种重要方法。
[Objective] To investigate the effects of survivin antisense oligodeoxy-nucleotid （ASODN） on proliferation and apoptosis in human colorectal carcinoma HT29 cells. [Methods] ASODN targeting survivin gene were transferred into HT29 cells by lipofectin reagent. MTT assay was used to measure the growth inhibitory rate and IC50, so as to observe the cytotoxicity of survivin ASODN on HT29 cells; The morphology changes of the nucleons were observed by Hoechst33342/PI staining. Flow cytometry （FCM） shows the cell cycle. [Results] After the HT29 cells were treated by different concentrations of survivin ASODN for 44 hours, the IC50 were 1 000 nmol/L, and the growth inhibitory rate in 200, 400, 600, 800 and 1 200 nmol/L group was （12.38 ± 7.96）%, （22.30 ± 4.49）%, （26.51 ± 3.08） %, （38.90 ± 3.66） %and （63.97 ± 4.50） %. When HT29 cells were treated with 1 000 nmol/L of survivin ASODN for 36 hours, distinct morphology changes of cell apoptosis were observed by Hoechst33342/PI staining. FCM shows the number of G2 phase cell were increased. [Conclusion] Survivin ASODN can effectively suppress the proliferation and induce apoptosis of human colorectal carcinoma HT29 cells. Survivin target therapy may be used as an important method in colorectal carcinoma treatment.
出处
《武警医学院学报》
CAS
2006年第6期517-520,F0002,共5页
Acta Academiae Medicinae CPAPF
