survivin反义寡核苷酸激活caspase-3诱导SGC7901胃癌细胞凋亡的实验研究  

Effect of survivin antisense oligonucleotide-activated caspase-3 on inducing apoptosis of human SGC7901 gastric cancer cells

作　　者：何金兰[1] 孔庆兖[1] 

机构地区：[1]徐州医学院病理学教研室,江苏徐州221002

出　　处：《徐州医学院学报》2009年第5期301-304,共4页Acta Academiae Medicinae Xuzhou

基　　金：江苏省教育厅自然科学研究计划(04KJB310143)

摘　　要：目的研究转染生存素(survivin)反义寡核苷酸诱导SGC7901胃癌细胞凋亡及其可能的作用机制。方法实验分为空白对照组(Control组)、脂质体组(Lip组)、正义寡核苷酸组(Sodn组)及反义寡核苷酸组(Asodn组)。人工合成正、反义寡核苷酸,经脂质体将survivin正、反义寡核苷酸转染入胃癌细胞48h后,MTT检测细胞增殖抑制率,Western blot检测caspase-3蛋白表达改变,Hoechst染色观察细胞凋亡形态改变及流式细胞仪检测凋亡率。结果Hoechst染色荧光显微镜下Control组、Lip组及Sodn组细胞核呈正常的蓝色,而Asodn组细胞核染色增强、核质浓缩、核碎裂,呈凋亡改变;Asodn组细胞增殖抑制率、细胞凋亡率、caspase-3蛋白表达均增高,与Control组、Lip组及Sodn组相比差异均有统计学意义(P<0.05)。结论survivin反义寡核苷酸可诱导胃癌细胞凋亡,其作用机制可能是通过激活caspase-3表达,启动凋亡信号传导。Objective To investigate the effect of transfected smvivin antisense oligonncleotide on inducing apoptosis in gastric cancer cells and the possible causative mechanism. Methods The experiment included four groups： control, liposome （Lip）, sense oligonucleotide （Sodn） and antisense oligonucleotide （Asodn）. Artificially synthesized survivin antisense oligonucleotide was transfected for 48 hours into gastric cancer cells using liposome. After cultivation, the cell proliferation inhibition rate was evaluated by MTT assay, the expression of caspase - 3 protein was determined with Western blot, and the morphological changes of apoptosis were observed by Hoechst staining, and the apoptosis rate was tested with Annexin v - FITC assay of flow cytometric analysis. Results Fluorescence microscopic observations revealed that Hoechst staining in the control, Lip and Sodn groups had nucleus presented as normal blue, while the Asodn group had such changes as reinforced nucleus staining, karyoplasm concentration, karyorrhexis and apoptotic modifications. Asodn group had a significant increase in the cell proliferation inhibition rate, cell apoptosis rate and expression of caspase - 3 protein, compared with the control, Lip and Sodn groups. The differences had statistical significance （ P 〈 0. 05 ）. Conclusion Survivin antisense oligonucleotide can induce apoptosis of gastric cancer cells, Its possible mechanism might be that the apoptosis signal transduction is initiated by the activation of caspase - 3 expression.

关 键 词：生存素 反义寡核苷酸 CASPASE-3 胃癌细胞 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]