微小RNA-155反义寡核苷酸对乳腺癌MDA-MB-157细胞及裸鼠移植瘤的作用  

Effects of antisense oligonucleotide targeting microRNA-155 on breast carcinoma cell line MDA-MB-157 and the implanted tumor in nude mice

作　　者：郑书荣[1] 郭贵龙[1] 黄奇迪[1] 黄督平[1] 尤捷[1] 黄关立[1] 

机构地区：[1]温州医科大学附属第一医院肿瘤外科,浙江温州325015

出　　处：《温州医学院学报》2014年第3期183-188,共6页Journal of Wenzhou Medical College

摘　　要：目的:探讨微小RNA-155(microRNA-155,miR-155)反义寡核苷酸(antisense oligonucleotide,ASO)对乳腺癌MDA-MB-157细胞增殖、凋亡及裸鼠移植瘤的作用。方法:设计合成化学修饰的miR-155 ASO,通过LipofectamineTM 2000转染MDA-MB-157细胞,激光共聚焦检测转染率,real-time PCR测定转染后miR-155表达水平的变化,CCK-8法检测细胞增殖抑制率,流式细胞仪检测细胞凋亡率,观察miR-155 ASO对裸鼠成瘤能力的影响,免疫组织化学法检测瘤体组织Caspase-3的表达。结果:激光共聚焦检测转染率达80%以上。转染miR-155 ASO后,miR-155表达明显下降,细胞增殖能力降低,凋亡增加。miR-155 ASO能明显抑制裸鼠移植瘤生长,抑制率为52.98%,并且能显著增加Caspase-3的表达。结论:miR-155 ASO能显著下调miR-155的表达水平,继而抑制乳腺癌MDA-MB-157细胞的增殖,促进其凋亡;并且通过增加靶基因Caspase-3的表达,抑制裸鼠移植瘤的生长,为miR-155作为乳腺癌治疗靶点提供了实验依据。To explore the effects of antisense oligonucleotide targeting miR-155 on proliferation and apoptosis in MDA-MB-157 cells and the growth of implanted tumor in nude mice. Methods： 2＇Ome modified antisense oligodeoxynucleotide targeting miR-155 was synthesized and then transfected into MDA- MB-157 cells by LipofectamineTM 2000. Transfection efficiency was detected by laser confocal microscope. Real- time RT-PCR method was used to detect the expression of miR-155. Cell counting Kit-8 （CCK-8） assay was used to detect the proliferation of MDA-MB-157 cells and cell apoptosis rate was measured by flow cytometry. The effects of miR-155 Antisense oligonucleotide （miR-155 ASO） on the transplanted tumor were assessed in nude mice. Results： Transfection efficiency detected by laser confocal microscope was higher than 80%. The level of miR-155 expression was significantly decreased （P〈0.05） in the cells transfected with miR-155 ASO, compared with that in cells transfected with negative control. After being transfected with miR^155 ASO, the viability of MDA-MB-157 cells reduced greatly （P〈0.05） and the number of apoptotic cells increased significantly. Additionally, miR-155 ASO inhibited the growth of transplanted tumor in vivo （inhibition ratio--52.98%） and significantly increased the expression of caspase-3. Conclusion： miR-155 ASO can effectively down-regulate the expression of miR-155, induce cell apoptosis and inhibit cell proliferation and tumor growth by increasing the expression of caspase-3. These findings provide the experimental evidence for using miR-155 as a therapeutic target of breast carcinoma.

关 键 词：乳腺肿瘤 微小RNA-155 反义寡核苷酸 MDA—MB 157细胞 裸鼠 

分 类 号：R73[医药卫生—肿瘤]