摘要
目的观察 Survivin ASODN 联合长春新碱(VCR)、5-氟尿嘧啶(5-Fu)对人结肠癌细胞系 HT-29凋亡的影响。方法设计合成特异性 Survivin 反义寡核苷酸(ASODN)。HT-29细胞分成6组:空白对照组、空脂质体转染对照组、正义链转染对照组、100、200、300 nmol/L 反义链转染组。以阳离子脂质体为载体转染至 HT-29细胞内,用 Western blot 法检测各组细胞 Survivin 蛋白表达情况;流式细胞仪检测细胞凋亡情况;MTT 法检测 VCR、5-Fu 对转染前后细胞的生长抑制情况。结果各浓度 ASODN 转染组癌细胞 Survivin 蛋白表达有不同程度减少,而各对照组细胞 Survivin 蛋白表达无明显变化。各 ASODN 转染组 VCR、5-Fu 对肿瘤细胞生长抑制率明显高于各对照组(P<0.05),且300 nmol/L 转染组明显高于100和200 nmol/L 组(P<0.05)。ASODN 转染组细胞凋亡指数明显高于各对照组(P<0.05),以300 nmol/L 转染组加 VCR 作用后凋亡指数最高(P<0.01)。结论 Survivin ASODN 转染人结肠癌细胞能下调 Survivin 蛋白的表达,诱导结肠癌细胞凋亡,抑制细胞增值,增加结肠癌细胞对化疗药物的敏感性。
Objective To investigate the effect of Survivin antisense oligonucleotide （ASODN） combined with Vinblastine （VCR） and 5-Fluorouracil （5-Fu） on the expression of Survivin mRNA and protein,and the apoptosis in colonic cancer cell lines. Methods The colonic cancer cell lines, HT-29 were transfected with Survivin ASODN （ 100,200,300 nmol/L） , SODN （ 100,200,300 nmol/L） , ODN （100,200,300 nmol/L） , by using Oligofectamine for 24 hour. The growth inhibition ratio （GI） in pre-and post-transfection cells treated by VCR and 5-Fu was detected by MTT method. The expression of Survivin protein was measured by western blotting assay, the apoptosis index （AI） and proliferation index were measured by flow cytometry （FCM）. Results Surviviti ASODN down-regulated the expression of the Survivin protein in HT-29 cell lines in dose-dependent manners, and at concentration of 300 nmol/L for 24 hour. But the expression of Survivin protein was not marked change in all control groups. By analyzing GI, it was demonstrated that GI was increased with dose-dependent manners in ASODN groups. Its maximum inhibition effect was achieved at a concentration of 300 nmoL/L. But GI at per control group was signifi- candy lower than ASODN groups （P 〈0.05 By analyzing the cell cycle and apoptosis, it was demonstrated that AI was increased and apoptosis was induced in per ASODN group. Its maximum effect was achieved at a concentration of 300 nmol/L and instantaneous treated by VCR. Conclusion Survivin ASODN can down-regulate the expression of Survivin protein when it was transfected into colonic cancer cells. Survivin ASODN also can induce apoptosis and suppress cell proliferation in dose-dependent manners, and increase the sensitivity of colonic cancer cells to chemotherapeutics.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2007年第12期1548-1550,共3页
Chinese Journal of Experimental Surgery
