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

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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。

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关键词 ：乳腺肿瘤, 微小RNA-155, 反义寡核苷酸, MDA-MB-157细胞, 裸鼠

Abstract：Objective: 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扥me 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.

Key words： breast neoplasms microRNA-155 antisense oligonucleotide MDA-MB-157 cells nude mice

收稿日期: 2013-03-19

通讯作者: 郭贵龙，主任医师，硕士生导师，Email：guogui long@sina.com。

作者简介: 郑书荣（1985-），男，浙江泰顺人，住院医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2014/V44/I3/183

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