生命科学研究2012，Vol.16Issue(2)：158-163,6.
沉默Bmi-1基因表达稳定细胞株的建立及其生物学特性的研究
Establishment and Biochemical Characteristics Study of Cell Line with Stable Silencing of Bmi-1 Gene
摘要
Abstract
B-cell-specific moloney murine leukemia virus insertion site 1 (Bmi-1) gene is a component of the Polycomb group, which is involved in regulation of cell proliferation. Study reveal that Bmi-1 may participate in tumorigenesis and may be used as a potential therapeutic target for the treatment of cancer. RNA interference (RNAi) was introduced to silence Bmi-1 gene aiming to study the effect on biochemical characteristics of MCF-7 such as invasion and metastasis and the role of Bmi-1 on breast cancer tumorigenesis. Retroviruses were produced using the retroviral vector and PT67 packaging cell line. Retroviruses were used to infect MCF-7 cell, then puromycin-resistant cells were pooled. A stable cell line with a persistent silencing of Bmi-1 was established, which was named as MCF-7/Bmi-lsi. The expression of Bmi-l was appraised with RT-PCR and Western blot assays, respectively. The colony-forming unit assay was used to measure the ability of cell proliferation, and Transwell chamber model was employed to test the ability of cell invasion and metastasis in vitro. Compared with MCF-7 and MCF-7/GFPsi, the mRNA and protein expression of Bmi-1 in MCF-7/Bmi-l were significantly less (P?0.05). The number and rate of colony formation in MCF-7/Bmi-lsi were also largely decreased (P?0.05). The results of invasion and metastasis assays indicated: the cells in MCF-7/ Bmi-l si moved from the upper chamber into the lower one for 24 h were less than those in the control groups MCF-7 and MCF-7/GFPsi (P?0.05). The stable cell line with a persistent silencing of Bmi-l gene was successfully established. Results reveal that the stable cell line with a persistent silencing of Bmi-l gene was successfully established. Silencing of Bmi-l gene can inhibit the ability of proliferation, invasion and metastasis in MCF-7 cells in vitro.关键词
Bmi-1基因/乳腺癌/RNA干扰/MCF-7细胞Key words
Bmi-l gene/ breast cancer/ RNA interference/ MCF-7 cell分类
医药卫生引用本文复制引用
武向梅,余华荣,卜友泉,易发平,汪长东,刘革力,苟小燕,邓小园,宋方洲..沉默Bmi-1基因表达稳定细胞株的建立及其生物学特性的研究[J].生命科学研究,2012,16(2):158-163,6.基金项目
重庆市自然科学基金资助项目(cstc2011jjA10035) （cstc2011jjA10035）
