Livin靶向ASODN对前列腺癌PC3细胞生物学特性的影响  

Effect of antisense oligonucleotide targeting Livin gene on biological characteristics of human prostate cancer PC3 cells

作　　者：谢晓强[1] 章振保[1] 杨恩明[1] 李显文[1] 李宗金[2] 徐勇[3] 

机构地区：[1]厦门市第二医院泌尿外科,福建厦门361021 [2]南开大学生物活性材料教育部重点实验室 [3]天津市泌尿外科研究所,天津医科大学第二医院泌尿外科

出　　处：《临床泌尿外科杂志》2013年第4期300-304,共5页Journal of Clinical Urology

基　　金：福建省自然科学基金资助项目(编号2008-59-14)

摘　　要：目的:合成Livin靶向的反义核苷酸(ASODN),观察其对前列腺癌PC3细胞凋亡和增殖等生物学特性的影响。方法:合成全硫代磷酸化修饰的Livin ASODN并通过脂质体转染前列腺癌PC3细胞。采用MTT法检测其对细胞增殖的影响,采用RT-PCR检测转染后Livin基因mRNA的表达情况,采用流式细胞仪检测转染后细胞的凋亡效应,采用体内成瘤实验比较细胞在裸鼠体内成瘤性的变化,采用激酶法测定Caspase-3活性的变化。结果:Livin ASODN转染PC3细胞后,与对照组相比,实验组细胞内Livin mRNA的表达水平下调(P<0.01);细胞的增殖受到显著抑制(P<0.01);细胞凋亡率明显增高(P<0.01);肿瘤细胞在荷瘤鼠体内的成瘤体积小于对照组(P<0.05);Caspase-3活性明显增加(P<0.05)。结论:靶向Livin基因的ASODN干扰了前列腺癌PC3细胞中Livin基因的表达,抑制了细胞的增殖并诱导其凋亡,延缓了肿瘤细胞的生长。Objective：To observe the effect of Livin on biology characteristics, such as apoptosis and proliferation, of human prostate cancer cells by antisense oligonucleotide targeting Livin gene. Method：Specific phosphorothioate antisense oligodeoxynucleotides targeting livin were synthesized and then transfected into PC3 cells. Proliferation was detected by MTT assay. The expressions of liven mRNA were detected by Real-time PCR. Cell apoptosis was assayed by flow cytometry. The in vivo tumor growth was observed in nude mice. The activity of Caspase-3 was detected by colorimetric assay. Result： After transfection, down-regulation of Livin mRNA expression in PC3 cells was found （P^0.01）. Compared with the control group, the experimental group showed an increase apoptosis rate （P^0.01）, a decreased ceil proliferation （P〈0.01）, an enhanced caspase-3 activity （P〈0. 05） and bigger tumor size in nude mice （P〈0.05）. Conclusion：The antisense oligonucleotide targeting Livin gene was constructed and can knockdown the expression of livin mRNA. It can inhibit PC3 cell proliferation, induce ap- omosis and inhibit tumor growth in vivo.

关 键 词：前列腺癌 LIVIN基因 反义寡核苷酸类 细胞凋亡 基因治疗 

分 类 号：R737.25[医药卫生—肿瘤]