[1]李海玉,宋方洲,卜友泉,等.过表达Bmi-1基因对乳腺癌细胞株BT474侵袭转移能力的影响[J].第三军医大学学报,2014,36(21):2187-2190.
 Li Haiyu,Song Fangzhou,Bu Youquan,et al.Effects of over-expression of Bmi-1 on invasion and metastasis in human mammary carcinoma cell line BT474[J].J Third Mil Med Univ,2014,36(21):2187-2190.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
36卷
期数:
2014年第21期
页码:
2187-2190
栏目:
论著
出版日期:
2014-11-15

文章信息/Info

Title:
Effects of over-expression of Bmi-1 on invasion and metastasis in human mammary carcinoma cell line BT474
作者:
李海玉宋方洲卜友泉易发平朱慧芳汪长刘革力武向梅
重庆医科大学基础医学院:分子医学与肿瘤研究中心,生物化学与分子生物学教研室,生理学教研室
Author(s):
Li Haiyu Song Fangzhou Bu Youquan Yi Faping Zhu Huifang Wang Changdong Liu Geli Wu Xiangmei

Chongqing Molecular Medical and Cancer Research Center, Department of Biochemistry and Molecular Biology, Department of Physiology, College of Basic Medical Sciences, Chongqing Medical University, Chongqing, 400016, China

关键词:
乳腺癌Bmi-1BT474过表达侵袭转移
Keywords:
breast cancer Bmi-1 BT-474 cells over-expression invasion and metastasis
分类号:
R394.2; R73-37; R737.9
文献标志码:
A
摘要:

目的      建立过表达Bmi-1的乳腺癌稳定细胞株,检测过表达Bmi-1对该肿瘤细胞株侵袭转移能力的影响,探讨Bmi-1与乳腺癌转移的相关性。      方法       根据GenBank中人源Bmi-1的mRNA序列,设计引物,构建Bmi-1真核表达载体GV144-Bmi-1,将其转染乳腺癌细胞株BT474,G418筛选建立过表达Bmi-1的稳定细胞株。实时荧光定量PCR和Western blot检测Bmi-1的mRNA和蛋白表达水平,Matrigel和Transwell侵袭小室模型检测转染和未转染BT474细胞体外侵袭和转移能力的变化,荧光定量PCR和Western blot检测细胞中MMP-2、MMP-9蛋白的变化。      结果       成功构建人源Bmi-1真核表达载体GV144-Bmi-1,转染BT474细胞后,获得稳定转染细胞株。实时荧光定量PCR和Western blot结果显示,与空载体组相比,实验组Bmi-1的mRNA和蛋白表达水平明显增高(P<0.05)。Bmi-1过表达后,迁移能力明显增强、穿膜细胞数明显增多,MMP-2、MM-9 mRNA和蛋白表达水平显著增强(P<0.05)。      结论       过表达Bmi-1稳定细胞株构建成功,Bmi-1表达上调可显著提高乳腺癌细胞的体外侵袭转移能力。

Abstract:

Objective       To construct a stable breast cancer cell line with over-expression of human Bmi-1 gene and investigate the effects of Bmi-1 over-expression on the invasion and metastasis in human breast cancer BT474 cells.       Methods       A pair of primers of Bmi-1 was designed according to the mRNA sequence in GenBank, and then an eukaryotic expression vector GV144-Bmi-1 was constructed. Human breast cancer BT474 cells were transfected with GV144-Bmi-1 and then treated with G418 to screen the cells with stable expression of Bmi-1. The mRNA and protein expression of Bmi-1 was detected by QRT-PCR and Western blotting. The invasion and metastatic ability of the BT474 cells with or without Bmi-1 over-expression were determined by Matrigel invasion assay and Transwell migration assay. The expression of MMP-2 and -9 was also detected at the mRNA and protein levels.       Results       The eukaryotic expression vector GV144-Bmi-1 was successfully constructed. A cell line with stable over-expression of Bmi-1 was obtained in BT474 cells. qRT-PCR and Western blotting indicated that the expression of Bmi-1 was significantly higher in the GV144-Bmi-1 over-expression group than in the control group (P<0.05). The cells with stable over-expression of Bmi-1 showed significantly enhanced migration ability with more cells moved from the upper chamber into the lower, and obviously increased expression of MMP-2 and -9 at the mRNA and protein levels (P<0.05).       Conclusion       A stable cell line with a persistent over-expression of Bmi-1 gene is established. Bmi-1 over-expression increases the ability of invasion and metastasis in human breast cancer BT474 cells.

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更新日期/Last Update: 2014-11-05